CN109517732B

CN109517732B - Integrated DNA analysis system

Info

Publication number: CN109517732B
Application number: CN201710852605.7A
Authority: CN
Inventors: 王承; 林小靖; 钱宇; 曹健荣
Original assignee: Denogen Beijing Bio Sci&tech Co ltd
Current assignee: Denogen Beijing Bio Sci&tech Co ltd
Priority date: 2017-09-19
Filing date: 2017-09-19
Publication date: 2022-12-16
Anticipated expiration: 2037-09-19
Also published as: CN109517732A

Abstract

The invention discloses an integrated DNA analysis system, which comprises: the reagent supply system comprises a reagent storage device and a reagent injection device; the microfluidic reaction system comprises a microfluidic chip and a microfluidic device, a sample to be analyzed can be added onto the microfluidic chip, a reagent injection device can inject a reaction reagent in the reagent storage device into the microfluidic chip, and the microfluidic device can control the flow of fluid in the microfluidic chip; the separation detection system comprises a capillary electrophoresis separation device and a fluorescence detection device, wherein reactants on the microfluidic chip can enter the capillary electrophoresis separation device, and the fluorescence detection device can detect a separation result in the capillary electrophoresis separation device. The integrated DNA analysis system has the characteristics of function integration, simplicity in operation, flexibility in use, rapidness in detection and mobile portability, and can be applied to large-scale commercial application.

Description

Integrated DNA analysis system

Technical Field

The invention relates to the field of DNA analysis and detection, in particular to an integrated DNA analysis system.

Background

DNA analysis techniques have a wide range of applications. In the field of basic research, DNA analysis is mainly applied to genome sequencing, gene expression profiling, gene mutation and polymorphism analysis, and the like; in the field of clinical medicine, the diagnosis of clinical diseases and the research and development of medicines can be carried out; in the field of judicial identification, individual identification and paternity identification can be carried out; in the agricultural field, animal and plant crossbreeding research and transgenic food safety detection can be carried out.

In the forensic field, for example, with the development of science and technology, the DNA detection technology of forensic has become an indispensable scientific tool for individual identification and crime fighting at the public security line. For decades, forensic DNA detection technology has undergone three technological revolution including multi-site DNA fingerprint analysis technology, amplified fragment length polymorphism analysis technology and mitochondrial DNA detection technology. At present, a fluorescence labeling multi-locus STR multiplex amplification detection technology, a mitochondrial DNA detection technology and an SNP analysis technology are developed as leading technical systems.

The realization of the above technologies does not leave the corresponding detection platform, and the development of the forensic DNA detection instrument determines the practical performance and the development speed of various forensic DNA detection technologies. Therefore, such instruments are becoming a research hotspot in the field of forensic science and in the field of analytical instruments. As an indispensable platform and tool for forensic DNA testing, mainstream forensic DNA testing instruments have undergone approximately three stages of development.

The first generation of DNA detection devices was generated in the 80's of the last century, and its core technology was slab gel electrophoresis, where the electrophoresis channel consisted of a solid medium containing many small pores and a buffer, DNA molecules were run through the gel, and after electrophoretic separation, the stained DNA analysis was fluorescence-excited and photographed using a UV light source and a camera. Early multi-site DNA fingerprint images, AMPFLP, STR alleles and the like are detected by adopting slab gel electrophoresis, and a large amount of technical personnel is required to participate due to low detection efficiency.

The second generation of DNA detection instrument is based on integration and automation, and begins in the 90 s of the last century, and lays a foundation for large-scale gene sequencing. In 1995, U.S. AB company introduced a 377 type plate-type genetic analyzer, which separated DNA molecules using a thin-layer polyacrylamide gel and obtained the sequence of a sample to be tested by reading these four fluorescent signals using Sanger's end-termination method, four-color fluorescent labels. The plate electrophoresis adopted by the instrument is troublesome in glue spreading, so that the instrument is quickly replaced by a platform adopting a capillary electrophoresis technology.

With the increasing demand for large-scale, high-throughput DNA analyzers, the third generation of automated fluorescence capillary gel electrophoresis analyzers has come into play. The novel instrument uses the capillary to replace the traditional vertical electrophoresis plate, overcomes the defect that manual glue making and manual lane identification are needed in the vertical plate electrophoresis, and leads the DNA analysis technology to completely move to the path of low cost, high flux, automation and scale. As the heat dissipation area of the unit volume of the capillary gel is much larger than that of the plate-type gel, the sensitivity and the resolution capability of DNA analysis are improved, and much higher voltage than that of plate-type gel electrophoresis can be used, so that the electrophoresis speed is improved by multiple times, the operation of vertical plate electrophoresis for 3 times a day is increased to 12 times a day, and the capillary gel becomes the mainstream technology which is most widely applied and has the mature technology at present. However, most of the existing devices have defects of different degrees, such as higher requirements on the specialty of operation, unsuitability for non-professionals, and difficulty in large-scale application and popularization in the conventional forensic DNA detection device in a short period.

Disclosure of Invention

The invention aims to provide an integrated DNA analysis system with functions integration, simple operation, flexible use, rapid detection and mobile portability.

In order to achieve the above purpose, the integrated DNA analysis system of the present invention has the following specific technical scheme:

an integrated DNA analysis system, comprising: a reagent supply system comprising a reagent storage device and a reagent injection device, the reagent storage device being in fluid communication with the reagent injection device; the microfluidic reaction system comprises a microfluidic chip and a microfluidic device, wherein a reagent injection device is in fluid communication with the microfluidic chip, the microfluidic chip is connected with the microfluidic device, a sample to be analyzed can be added onto the microfluidic chip, a reaction reagent in the reagent storage device can be injected into the microfluidic chip by the reagent injection device, and the microfluidic device can control the flow of fluid in the microfluidic chip; the separation detection system comprises a capillary electrophoresis separation device and a fluorescence detection device, wherein the microfluidic chip is in fluid communication with the capillary electrophoresis separation device, the fluorescence detection device is connected with the capillary electrophoresis separation device, reactants on the microfluidic chip can enter the capillary electrophoresis separation device, and the fluorescence detection device can detect a separation result in the capillary electrophoresis separation device.

Further, the reagent storage device comprises a conventional reagent kit and a temperature control reagent kit, wherein the conventional reagent kit comprises a reagent bottle and can store reagents with low requirements on the environmental temperature; the temperature control kit comprises a reagent bottle and a temperature control device, wherein the temperature control device can adjust the temperature of the reagent in the reagent bottle and can store the reagent with high requirement on the environmental temperature.

Further, the temperature control kit comprises a heat preservation sheath, the reagent bottle is arranged in the heat preservation sheath, the temperature control assembly is arranged on the heat preservation sheath, and the reagent bottle, the heat preservation sheath and the temperature control assembly are arranged in the protection box body.

Further, the reagent injection device comprises a reagent accommodating chamber and an automatic pushing device, a reagent inlet of the reagent accommodating chamber is communicated with a reagent outlet of the reagent storage device in a fluid mode, and a reagent outlet of the reagent accommodating chamber is communicated with a reagent inlet of the microfluidic chip in a fluid mode; the automatic pushing device is connected with the reagent containing chamber, and can add the reagent in the reagent storage device into the reagent containing chamber and fill the reagent in the reagent containing chamber onto the microfluidic chip.

Furthermore, the reagent injection device comprises a plurality of injectors which are vertically arranged side by side, each injector is connected with a reagent push rod, each reagent push rod is respectively connected with a power assembly, and the power assemblies can respectively drive the pushing assemblies to reciprocate in the injectors so as to inject the reagents into the microfluidic chip in a classified mode.

Further, the microfluidic chip comprises a chip body and a reaction unit, wherein the chip body is provided with a sample extraction unit and a fluid pipeline, and the sample extraction unit is used for extracting reactants; the reaction unit is attached to the chip body and provided with a reaction area, the reaction area on the reaction unit is communicated with the fluid pipeline on the chip body, and reactants and reaction reagents can be conveyed into the reaction area through the fluid pipeline so as to complete reaction in the reaction area.

Further, the chip body comprises an accessory piece unit, a pipeline piece unit and a control valve piece unit which are arranged in an overlapped mode, a sample extraction accessory is arranged on the accessory piece unit, a fluid pipeline is arranged on the pipeline piece unit, and a control valve piece for controlling the on-off of the fluid pipeline is arranged on the control valve piece unit.

Furthermore, a pipeline disconnection point is arranged on the fluid pipeline of the pipeline sheet unit, a pipeline connection hole is arranged at the position, corresponding to the pipeline disconnection point, of the control valve sheet unit, the pipeline connection hole is communicated with the fluid pipeline near the pipeline disconnection point, and the fluid pipeline on the pipeline sheet unit is communicated at the pipeline disconnection point through the pipeline connection hole.

Furthermore, the opening and closing valve is arranged at the position, corresponding to the pipeline connecting hole, on the control valve plate unit, and the opening and closing valve can control the opening and closing of the pipeline connecting hole so as to realize the communication and disconnection of the corresponding fluid pipeline on the pipeline plate unit.

Furthermore, the micro-fluidic device comprises a chip bearing component and a connecting hole opening and closing component, the micro-fluidic chip can be placed on the chip bearing component, the connecting hole opening and closing component is connected with a pipeline connecting hole in a chip placed on the chip bearing component, and the opening and closing of the pipeline connecting hole in the chip can be controlled so as to realize the connection and disconnection of a fluid pipeline connected with the pipeline connecting hole on the chip.

Furthermore, a temperature control assembly is arranged on the chip bearing assembly, and after the micro-fluidic chip is placed on the chip bearing assembly, the temperature control assembly is in contact with the micro-fluidic chip, so that the reaction temperature on the micro-fluidic chip can be adjusted.

Furthermore, the micro-fluidic device comprises a mechanical control assembly, wherein the mechanical control assembly is connected with the chip bearing assembly and can drive the chip bearing assembly to move in a reciprocating manner.

Further, the capillary electrophoresis separating device comprises a capillary electrophoresis component and a temperature control component, the capillary electrophoresis component comprises a protection component, and the capillary is arranged in the protection component; the temperature control assembly comprises an isolation assembly, and the heating assembly is arranged inside the isolation assembly; the capillary electrophoresis component and the temperature control component are in contact with each other and can conduct heat transfer, and the heating component in the temperature control component can conduct temperature adjustment on the capillary in the capillary electrophoresis component.

Furthermore, a detection window is arranged at a position, corresponding to the fluid outlet end of the capillary tube, on the protection assembly, and the electrophoresis separation in the capillary tube can be detected through the detection window.

Furthermore, the capillary electrophoresis separating device comprises a quick locking component, and the quick locking component is connected with the capillary electrophoresis component so as to detachably arrange the capillary electrophoresis component in the integrated DNA analysis system.

Further, the capillary electrophoresis separation device comprises a gel pushing device, the gel pushing device comprises a gel containing chamber, a temperature adjusting assembly and an automatic pushing device, the gel containing chamber is used for temporarily storing gel, the temperature adjusting assembly is connected with the gel containing chamber and can adjust the temperature of the gel in the gel containing chamber, the automatic pushing device is connected with the gel containing chamber and can add the gel into the gel containing chamber and push the gel in the gel containing chamber into the capillary electrophoresis assembly.

Furthermore, the gel propelling device comprises a reagent cylinder and a reagent push rod, a protection box is arranged on the outer side of the reagent cylinder, a semiconductor refrigerator is arranged in the protection box, the semiconductor refrigerator is connected with the reagent cylinder and can adjust the temperature of gel in the reagent cylinder, the reagent push rod is connected with the propelling assembly, and the propelling assembly can drive the reagent push rod to reciprocate in the reagent cylinder.

Further, the fluorescence detection device comprises a laser, an objective lens and a spectrometer, wherein the laser, the objective lens and the spectrometer are sequentially connected through an optical path, a first optical path adjuster is arranged between the laser and a capillary tube in the capillary tube electrophoresis separation device and can change a path of laser emitted by the laser, a second optical path adjuster is arranged between the objective lens and the spectrometer and can change a path of fluorescence between the objective lens and the spectrometer.

Further, the first optical path adjuster includes a first plane mirror and a first lens, the first plane mirror can vertically reflect the laser emitted by the laser to the capillary direction, and the first lens can focus the laser reflected to the capillary direction.

Further, the second light path adjuster comprises a second plane reflector, a filter and a second lens, wherein the second plane reflector can vertically reflect the fluorescence emitted by the capillary tube to the spectrometer; the filter can change the fluorescence reflected towards the spectrometer into a single color; the second lens may focus the monochromatic fluorescent light.

The integrated DNA analysis system of the invention has the following advantages:

1) All steps of DNA detection and analysis are integrated on a miniaturized device, the operation is convenient, the detection is rapid, the efficiency of DNA detection is greatly improved on the premise of ensuring the accuracy, meanwhile, the portability of movement is greatly improved by the integrated miniaturized device, and the range of applicable places is enlarged.

2) The unique split chip design reduces the whole manufacturing difficulty and cost and improves the reaction effect on the premise of realizing the integrated function.

3) The original micro-fluidic device realizes the fine control of the flow of the reaction fluid on the chip by using the fluid pipeline and the control valve which are designed in a layered way in a matching way, and ensures the smooth proceeding of the reaction on the chip.

4) The chip reaction system and the reagent supply system which are designed independently can meet the detection requirements of different quantities, and the flexibility of equipment use is improved.

5) The integrated capillary electrophoresis system reduces the difficulty of replacing the capillary, improves the reaction efficiency and is convenient for non-professional personnel to use in daily life.

6) From the sample to the result, a user does not need to operate in a standard PCR laboratory, only needs to carry out simple reagent preparation and sample adding processes, and the subsequent DNA extraction process, the PCR product mixing, the capillary electrophoresis process, the signal detection process and the like can be carried out automatically.

Drawings

FIG. 1 is a schematic structural view of an embodiment of the integrated DNA analysis system of the present invention;

FIG. 2 is a front view of the integrated DNA analysis system of FIG. 1;

FIG. 3 is a left side view of the integrated DNA analysis system of FIG. 1;

FIG. 4 is a right side view of the integrated DNA analysis system of FIG. 1;

FIG. 5 is a rear view of the integrated DNA analysis system of FIG. 1;

FIG. 6 is a schematic diagram of the structure of the microfluidic chip in the integrated DNA analysis system of the present invention;

FIG. 7 is a perspective view of the tubing of the microfluidic chip of FIG. 6;

FIG. 8 is an exploded view of the microfluidic chip of FIG. 6;

FIG. 9 is a schematic view of the top sheet of FIG. 8;

FIG. 10 is a schematic view of the construction of the accessory strip of FIG. 8;

FIG. 11 is a schematic view of the duct piece of FIG. 8;

FIG. 12 is a schematic view of the channel sheet of FIG. 8;

FIG. 13 is a schematic view of the valve plate of FIG. 8;

FIG. 14 is a schematic view of the structure of the backsheet of FIG. 8;

FIG. 15 is a schematic structural view of the reaction plate of FIG. 8;

FIG. 16 is a schematic view showing the structure of a microfluidic device in the integrated DNA analysis system of the present invention;

fig. 17 is an exploded view of the microfluidic device of fig. 16;

FIG. 18 is a schematic illustration of the mechanical control platform of FIG. 17;

FIG. 19 is a schematic structural view showing a first example of a reagent storage apparatus in the integrated DNA analysis system of the present invention;

FIG. 20 is a schematic structural view showing a second example of a reagent storage apparatus in the integrated DNA analysis system of the present invention;

FIG. 21 is a disassembled view of the reagent storage device of FIG. 20;

FIG. 22 is a schematic view showing the construction of a reagent injection device in the integrated DNA analysis system of the present invention;

FIG. 23 is a schematic view of the reagent injector of FIG. 22 with a panel omitted;

FIG. 24 is a schematic structural view of the reagent interface device of FIG. 22;

FIG. 25 is a schematic structural view of the slide assembly of FIG. 22;

FIG. 26 is a schematic diagram showing the structure of a capillary electrophoresis separation system in the integrated DNA analysis system of the present invention;

FIG. 27 is a schematic diagram of the structure of the capillary electrophoresis cartridge of FIG. 26;

FIG. 28 is a disassembled view of the capillary electrophoresis cartridge of FIG. 27;

FIG. 29 is a schematic diagram of the capillary heating cartridge of FIG. 26;

FIG. 30 is an exploded view of the capillary heating cartridge of FIG. 29;

fig. 31 is a schematic structural view of the card type electromagnetic locking device of fig. 26;

fig. 32 is an exploded view of the card-type electromagnetic locking device of fig. 31;

FIG. 33 is a schematic view showing the structure of a gel propelling system in the integrated DNA analyzing system of the present invention;

FIG. 34 is a side view of the gel self-advancing system of FIG. 33;

FIG. 35 is an exploded view of the gel self-advancing system of FIG. 33;

FIG. 36 is a schematic view showing the structure of a fluorescence detection system in the integrated DNA analysis system of the present invention.

Detailed Description

In order to better understand the purpose, structure and function of the present invention, an integrated DNA analysis system of the present invention will be described in detail with reference to the accompanying drawings.

Compared with the traditional DNA analysis equipment, the integrated DNA analysis system integrates almost all steps in the DNA analysis processes of sample extraction, PCR reaction, electrophoretic separation, fluorescence detection and the like, so that the DNA analysis process can be completed in the whole equipment, the use scene of the equipment is greatly improved, and the integrated DNA analysis system can be widely applied to the fields of public security, justice, clinic and the like.

According to the functional area division, the integrated DNA analysis system comprises a reagent supply system, a microfluidic reaction system and a separation detection system. The reagent supply system is used for storing various reagents required in the integrated DNA analysis system and selectively delivering the reagents to each part according to the requirements so as to meet the requirements of equipment operation; the microfluidic reaction system is mainly used for processing samples, such as DNA extraction, PCR reaction and the like, and preparing for subsequent separation and detection; the separation detection system is used for carrying out electrophoretic separation on the DNA fragments and carrying out fluorescence detection so as to meet the detection requirement. In addition, it should be noted that the integrated DNA analysis system of the present invention is also correspondingly provided with auxiliary structures such as a control system, a power system, etc. to achieve automation of the operation of the apparatus.

Microfluidic reaction system

The microfluidic reaction system comprises a microfluidic chip and a microfluidic device, wherein the microfluidic chip is an integrated chip, and various operations and functions of DNA analysis, such as DNA capture and washing, DNA amplification and the like, can be integrated on the microfluidic chip; the microfluidic device is used for controlling the fluid flow in the microfluidic chip so as to assist the microfluidic chip to realize the integrated operation.

Specifically, the microfluidic chip can integrate a plurality of basic operation units for sample preparation, reaction, separation, detection, and the like in the processes of biological, chemical, and medical analysis theoretically, but due to the limitation of technical development, too many operation steps are integrated on one microfluidic chip, so that the accuracy and precision of an experimental result are reduced, the use effect of the whole chip is greatly reduced, and the microfluidic chip can only be in the stage of laboratory research and cannot be applied to large-scale commercial application. The micro-fluidic chip only integrates two step units of DNA extraction and PCR reaction, and can ensure good use effect on the premise of realizing small integration.

Compared with the existing integral microfluidic chip, the microfluidic chip provided by the invention comprises two parts, namely a chip body and a reaction unit. The chip body is a main body part of the micro-fluidic chip and is integrated with a sample extraction unit, a fluid pipeline and the like; the reaction unit is a dedicated reaction area for carrying out the PCR reaction step. The chip body and the reaction unit are two parts which are independently designed, the materials and the shapes can be flexibly designed according to the requirements, and the formed chip body and the reaction unit are combined together to form the micro-fluidic chip.

The design considers that most biochemical reactions have special requirements on the material and the shape of the reaction container, and the special material cannot be used for manufacturing the chip body in a large scale or the special shape is not convenient to manufacture on the conventional chip body, so that the problems of high processing difficulty, high material cost, poor use effect and the like can be caused. Taking a PCR reaction as an example, a reaction container made of PP (Polypropylene) can obviously improve the reaction effect, but compared with the conventional chip material, the PP material has the problems of high processing difficulty, high material cost and the like, and is not suitable for being applied to the whole microfluidic chip.

In addition, the independent design of the reaction unit can also make the shape of the reaction region more flexible without considering the problems of the processing complexity of the chip body and the like, because a plurality of auxiliary structures such as fluid pipelines and the like are inevitably designed on the chip body. The reaction shape can be conveniently manufactured on the reaction unit in the invention, and because the reaction unit is almost provided with no additional auxiliary structure, the complexity of the manufacturing process is greatly reduced.

Meanwhile, the independently designed reaction unit is convenient for accurate temperature control, because the requirement of the PCR reaction in the microfluidic chip on the temperature is stricter, and the existing reaction unit integrated on the chip body is not ideal in temperature control.

In order to further reduce the manufacturing difficulty, the chip body of the present invention may be formed by stacking a plurality of sheet units, for example, a top sheet unit, an accessory sheet unit, a pipeline sheet unit, a channel sheet unit, a valve hole sheet unit, and a bottom sheet unit. Therefore, the complex structure of the chip body can be decomposed and realized by different sheet body units, such as manufacturing installation spaces of all the accessory units on the top sheet unit and the accessory sheet unit, manufacturing basic fluid pipelines on the pipeline sheet unit and the channel sheet unit, and manufacturing fluid pipeline control valves on the valve orifice unit and the bottom sheet unit, thereby facilitating the standardized and batch production of factories. The chip with the complex structure can be manufactured by uniformly performing superposition processing on the sheet body units manufactured in batches, and the superposition processing modes of the sheet body units can be various, such as adhesion, bonding and the like.

The chip body of the invention is mainly integrated with a sample extraction unit, a fluid pipeline unit and a waste liquid discharge unit. The sample extraction unit can extract a sample, the fluid pipeline unit can realize the flow of reaction fluid on the chip body, and the waste liquid discharge unit can absorb redundant reaction fluid on the chip body.

Taking DNA extraction as an example, the sample extraction unit can adopt a plurality of extraction methods, such as FTA test paper extraction, which is a proprietary technology of Whatman company, and is originally applied to DNA and RNA collection, transportation, purification and storage at room temperature, all the work is completed on one card, and is very suitable for being applied to miniaturized and integrated microfluidic chips. In addition, the existing magnetic bead method can also be used for extraction, and the magnetic bead method is a novel nucleic acid extraction technology taking nano biological magnetic beads as carriers, nucleic acid molecules can be specifically identified and combined with silicon hydroxyl on the surfaces of the magnetic beads and are gathered or dispersed under the action of an external magnetic field, so that the manual operation processes of centrifugation, supernatant extraction and the like in the traditional nucleic acid extraction process are thoroughly eliminated, and the automatic extraction of nucleic acid is realized. Therefore, the sample extraction unit on the microfluidic chip can adopt any mode capable of automatically extracting samples so as to meet the requirements of integration and automation.

The fluid pipeline unit is mainly used for connecting all the components on the microfluidic chip so as to realize the flow of the sample and the reagent among the components. The fluid pipeline unit mainly includes two aspects, namely, the driving of the fluid, namely how to drive the sample and the reagent to flow in the fluid pipeline, and the controlling of the fluid, namely, how to control the flow direction of the sample and the reagent in the fluid pipeline.

The micro-fluidic chip mainly adopts the micro-pump to realize the driving of the fluid, wherein the micro-pump can adopt a mechanical micro-pump, such as a centrifugal force micro-pump, a thermal power micro-pump, an electrostatic micro-pump, a pneumatic power micro-pump, an electromagnetic micro-pump, a piezoelectric micro-pump, a bimetallic memory alloy micro-pump and the like, the mechanical micro-pump can provide low-flow fluid conveying matched with a fluid channel on the micro-fluidic chip, and the micro-fluidic chip is particularly suitable for the simple interface assembly of high polymer material chips. Of course, according to practical situations, a non-mechanical micropump may be used to drive the flow on the microfluidic chip, such as an electric field force driven pump, a capillary micropump, a biological action micropump, a magnetohydrodynamic pump, an optical driven pump, a gravity-based driven pump, a chemical action micropump, and the like.

The invention realizes the control of fluid flow on the microfluidic chip by the special design of the fluid pipeline and the structure of the micro valve matched with the fluid pipeline. The chip body is provided with a plurality of fluid pipelines, and each fluid pipeline is communicated with a certain operation or reaction area on the microfluidic chip, so that a fluid reagent can flow to the operation or reaction area through the fluid pipeline. The chip body is provided with a flow connecting hole between the two corresponding fluid pipelines, and the connection or disconnection of the two fluid pipelines can be realized by opening or closing the flow connecting hole.

The opening or closing of the flow connection hole can be realized in various ways, for example, an elastic structure layer is arranged on one side of the flow connection hole, and an external force is applied to a position, corresponding to a certain flow connection hole, on the elastic structure layer, so that the elastic structure layer can seal the flow connection hole, the closing of the flow connection hole is realized, correspondingly, the two fluid pipelines are in an open state at the flow connection hole, and a fluid reagent cannot flow between the two fluid pipelines; when the external force is removed, the elastic structure layer can be automatically reset, the flow connecting hole is opened, correspondingly, the two fluid pipelines can be in a communication state through the flow connecting hole, and the fluid reagent can flow between the two fluid pipelines. Wherein the external force applied to the elastic structure layer can be realized by a solenoid valve, a pneumatic piston and the like.

Based on the above description, the microfluidic device of the present invention includes a chip carrying assembly and a connection hole opening and closing assembly, wherein the chip carrying assembly is used for carrying the microfluidic chip and providing a stable platform for operations and reactions on the microfluidic chip, and the connection hole opening and closing assembly is connected to the microfluidic chip, or specifically to a flow connection hole on the microfluidic chip, and is used for controlling the opening and closing of the flow connection hole on the microfluidic chip, so as to control the flow of fluid on the microfluidic chip.

The chip bearing component is preferably of a platform structure, the main function is to bear a chip, the chip is required to be accurately connected with the connecting hole opening and closing component after being loaded to the chip bearing component, so that a corresponding connecting groove or connecting block is arranged on the chip bearing component, and the chip can be directly placed on the connecting groove or connecting block so as to ensure accurate connection of the follow-up chip and the connecting hole opening and closing component. Based on the conventional loading habit of the chip, the chip bearing component is preferably provided with the corresponding connecting groove, so that the accuracy of the chip mounting position can be ensured, and the stability and the space saving during the subsequent operation of the chip can also be ensured.

The chip bearing component is generally horizontally arranged, the connecting groove arranged on the chip bearing component is also of a horizontal structure, and the chip is basically horizontally arranged after being loaded into the connecting groove, so that the relative independence among all areas on the chip can be ensured, and the influence of the gravity action on the operation and the reaction is avoided. However, depending on different situations, the chip carrier assembly may be arranged vertically or obliquely, and most of the chips may also be arranged vertically or obliquely, in which case the reliability of the chip loading in the connecting grooves is particularly important.

Can be provided with chip fixed knot on the chip loads the subassembly and construct, if only directly place the chip in the spread groove, can appear rocking the condition such as usually, it is bigger to the higher micro-fluidic system of requirement that becomes more meticulous influence, and chip fixed knot constructs in order to carry out extra fixed to the chip of loading in the spread groove, guarantees the stability after the chip loads. The chip fixing structure is preferably implemented by a simple structure, for example, a rotating clamping plate is arranged beside the connecting groove to directly and firmly clamp the chip in the connecting groove, or further to save space, an elastic buckle can be arranged inside the connecting groove, and when the chip is loaded into the connecting groove, the chip is directly buckled in the connecting groove through the elastic buckle. Because the fixing structure is more, the chip can be selectively used as long as the chip can be fixed, the structure is simple, and the space is saved.

The chip is fixed on the chip loading assembly and then needs to be connected with the connecting hole opening and closing assembly, and the connecting hole opening and closing assembly can adopt a solenoid valve, and the solenoid valve has higher sensitivity and mature technology and is very suitable for being applied to the field of microfluidics. Specifically, the plunger of the solenoid valve is inserted into the flow connection hole on the chip, and the opening and closing of the flow connection hole are controlled by the expansion and contraction of the plunger, for example, when the plunger is inserted into the flow connection hole, the flow connection hole is in a closed state, correspondingly, the two flow pipelines are in a disconnected state at the flow connection hole, and the fluid reagent cannot flow between the two flow pipelines; when the plunger is moved out of the flow connecting hole, the flow connecting hole is in an open state, the two corresponding flow pipelines are in a communication state through the flow connecting hole, and the fluid reagent can flow between the two flow pipelines.

Alternatively, an elastic structure layer may be further disposed between the flow connection hole on the chip and the plunger of the solenoid valve, so that the flow connection hole is opened and closed by the elastic structure layer. The method specifically comprises the following steps: when the plunger piston is close to the elastic structure layer, the position, opposite to the flow connecting hole, on the elastic structure layer is extruded, the elastic structure layer deforms, and then the flow connecting hole is blocked, so that the flow connecting hole is in a closed state, correspondingly, the two flow pipelines are in a disconnected state at the flow connecting hole, and a fluid reagent cannot flow between the two flow pipelines; when the plunger piston is far away from the elastic structure layer, the elastic structure layer can be restored to the initial state, the flow connecting hole is in an open state, correspondingly, the two flow pipelines can be in a communication state through the flow connecting hole, and the fluid reagent can flow between the two flow pipelines.

The connection hole opening and closing assembly is described above by taking a solenoid valve as an example, and the invention can also be realized by adopting other structures, such as a hydraulic or pneumatic piston and the like, as long as the effect of opening or closing the flow connection hole on the chip can be achieved.

The connecting hole opening and closing assembly can be selectively arranged relative to the position of the chip loading assembly, and two groups of arrangement modes are generally adopted, namely the connecting hole opening and closing assembly is positioned below a connecting groove on the chip loading assembly, and after a chip is loaded into the connecting groove, the connecting hole opening and closing assembly is inserted into a flowing connecting hole on the chip from the bottom of the chip; and the connecting hole opening and closing component is inserted into the flow connecting hole on the chip from the top of the chip after the chip is loaded into the connecting groove. As to which way to choose, it is generally based on the specific form of the chip, and if the flow connection hole of the chip needs to be inserted from the bottom, the first way is chosen, and if it needs to be inserted from the top, the second way is chosen. The overall structural arrangement of the microfluidic device is also considered, such as whether the chip loading assembly and the connection hole opening and closing assembly are fixedly connected together or are arranged relatively independently, the size of the upper space or the lower space of the chip loading assembly, and the like.

The solenoid valve connecting hole opening and closing assembly usually generates certain heat when acting, and the inside of the microfluidic device is relatively closed, so that the normal operation of each internal assembly is guaranteed, particularly the solenoid valve, the microfluidic device can be further provided with a heat dissipation assembly which can dissipate heat of internal components of the microfluidic device, and the heat dissipation mode preferably adopts airflow heat dissipation, so that the solenoid valve connecting hole opening and closing assembly has the advantages of simple structure and easiness in realization, and is relatively suitable for heat dissipation of the solenoid valve. However, if the opening and closing of the flow connection hole on the chip are realized by adopting a mechanical structure of other forms, a heat dissipation mode suitable for the mechanical structure, such as water cooling heat dissipation, may have a better effect than air flow heat dissipation.

After the fluid reagent on the control chip flows to the related reaction area through the connecting hole opening and closing assembly, the related reaction can be carried out, for example, after the DNA extraction is finished, the PCR reaction reagent and the extracted DNA flow to the PCR reaction area together, and then the PCR reaction can be started. Because the reaction on the chip mostly has specific requirements on the temperature, in order to ensure the normal operation of the reaction on the chip, a temperature control component is also arranged in the microfluidic device, and the temperature of the reaction on the chip can be adjusted through the temperature control component.

Wherein, the setting position of the temperature control assembly is determined based on the position of the reaction area on the chip so as to ensure the accurate control of the reaction temperature on the chip. For example, if the reaction region is located at the end of the chip, the temperature control member should be correspondingly disposed at a position opposite to the end of the chip, and if the reaction region is located at the middle of the chip, the temperature control member should be correspondingly disposed at a position opposite to the middle of the chip, and preferably, the temperature control member is in direct contact with the chip to ensure the accuracy of heat conduction.

Since the chip is fixedly arranged in the connecting groove of the chip loading assembly in the invention, the temperature control assembly is arranged opposite to the connecting groove, if the reaction region on the chip is positioned at the top, the temperature control assembly can be arranged above the connecting groove so as to facilitate the contact of the reaction region at the top of the chip, and if the reaction region on the chip is positioned at the bottom, the temperature control assembly can be arranged below the connecting groove or directly arranged on the bottom surface of the connecting groove so as to facilitate the contact of the reaction region at the bottom of the chip. The temperature control assembly is preferably a semiconductor refrigerator, the semiconductor refrigerator (Thermo Electric Cooler) is made of semiconductor materials by the Peltier effect, and has the characteristics of small volume, low energy consumption and high temperature change rate, and the refrigeration, heating and the refrigeration and heating rates can be determined by the direction and the magnitude of current, so that the temperature control assembly is convenient to use.

In order to facilitate chip replacement or other maintenance, a mechanical control assembly can be arranged, and the mechanical control assembly can move each assembly in the microfluidic device so as to move out of the whole equipment for convenient operation. Wherein, because need change the chip, so remove comparatively frequent mainly that the chip loads the subassembly, the mechanical control subassembly can be controlled the chip loading subassembly alone, when the chip is changed to needs, moves out the chip loading subassembly alone from whole equipment, simultaneously, for the maintenance of other parts of convenience, like the connecting hole opens and close subassembly, temperature control subassembly or radiator unit, also can be to its individual control or combination control.

In view of the integrity and stability of the microfluidic device, the chip loading assembly, the connection hole opening and closing assembly, the temperature control assembly and the heat dissipation assembly are generally fixed together, so the mechanical control assembly generally performs displacement control on the whole microfluidic device. The displacement control mainly comprises transverse displacement control and longitudinal displacement control, the longitudinal displacement control is used for adjusting the height of each component, each component is conveniently separated from or combined with the whole equipment, and the transverse displacement control is used for horizontally moving each component out of the whole equipment so as to replace chips or perform other maintenance.

The mechanical control assembly is generally in a structure form and generally comprises a power mechanism, a sliding mechanism and the like, each assembly of the microfluidic device can be fixedly arranged in the sliding mechanism, and the power mechanism drives the sliding mechanism to move so as to drive each assembly to move transversely or longitudinally. Because the whole apparatus is generally square, and generally placed on a plane when in use, and corresponding components, such as reagent conduits, are disposed above the chip loading assemblies in the whole apparatus, each assembly is generally moved longitudinally to be separated from the relevant assembly on the whole apparatus, and then is considered to be moved laterally out of the whole apparatus to perform operations such as chip replacement.

Reagent supply system

The reagent supply system comprises a reagent storage device and a reagent injection device, wherein the reagent storage device is used for temporarily classifying and storing reagents required by the equipment, and the reagent injection device is used for communicating the reagent storage device with corresponding equipment components so as to inject the reagents into the relevant equipment components according to the requirements, so that the reaction can be smoothly carried out.

Specifically, the reagent storage device includes two types, one is a conventional reagent storage case for temporarily storing a reagent, such as a buffer solution, etc., which requires less external temperature; the other is a reagent storage box with temperature control, which is used for temporarily storing reagents with high requirements on external temperature, such as DNA polymerase and the like. Among them, as for the first conventional reagent storage cassette, a reagent storage cassette common in the art may be employed, generally including only reagent bottles, tubes, etc.; for the second temperature control reagent storage box, a temperature control device can be added on the basis of a common reagent storage box, for example, a heat preservation sheath, a semiconductor refrigerator and the like are additionally arranged outside a reagent bottle so as to meet the requirement on temperature.

The reagent injection device is used for connecting the reagent storage device and the microfluidic chip, and comprises a reagent accommodating chamber, wherein the reagent accommodating chamber is used for temporarily storing a fluid reagent to be filled on the chip. However, other forms such as a kit can be used, which can be used to pump the reagents of the kit to the chip, and of course, there are many other forms, such as a syringe or a syringe, which are commonly used in practice.

The reagent inlet of the reagent holding chamber may be in fluid communication with a reagent storage device, and a fluid reagent in the reagent storage device may be pre-filled into the reagent holding chamber and then subsequently filled from the reagent holding chamber onto the chip. The control valve is generally arranged between the reagent accommodating chamber and the reagent storage device so as to control the delivery of the reagent to the reagent accommodating chamber according to the requirement, and the electromagnetic valve is preferably adopted for controlling in consideration of the quantity of the reagent required by the microfluidic chip, so that the fine control of reagent filling can be ensured. Thus, when it is desired to add reagent to the reagent-containing chambers, the corresponding solenoid valve is opened and reagent can be delivered from the reagent storage device to the reagent-containing chambers.

After the reagent is added into the reagent accommodating chamber, corresponding reagent can be filled onto the chip according to the reaction requirement on the chip, wherein a reagent outlet of the reagent accommodating chamber is communicated with a reagent inlet on the chip in a fluid manner, a control valve is generally arranged between the reagent accommodating chamber and the chip so as to control the delivery of the reagent onto the chip according to the requirement, and preferably, an electromagnetic valve is adopted so as to ensure the fine control of the reagent filling. Therefore, when the reagent in the reagent containing chamber needs to be filled on the chip, the corresponding electromagnetic valve is opened, and the reagent can be conveyed from the reagent containing chamber to the chip.

Since the reagent holding chamber is usually in the form of a syringe, when it is necessary to fill the reagent in the syringe onto the chip, the reagent in the syringe can be pushed out by pushing the push rod of the syringe, and the push rod descends. In consideration of the requirement of automation, the push rod of the injector is connected with the power assembly, and the power assembly can automatically drive the push rod to move downwards according to the preset condition. The power assembly generally adopts a combination form of a motor and a sliding block, the sliding block is fixedly connected with a push rod of the injector, and the motor can drive the sliding block to slide on a corresponding sliding rail, so that the push rod of the injector is driven to move upwards or downwards to push the reagent out of the injector. If a combination of a reagent kit and a delivery pump is used, the time and amount of reagents to be dispensed onto the chip are directly controlled by the delivery pump, and a syringe is preferably used for cost reasons.

Since it is preferable to control the reagent filling by driving the push rod with a power assembly, the movement distance of the push rod should be controlled to avoid excessive movement of the push rod, which may damage the syringe or the push rod, since the microfluidic chip determines that the syringe has a small volume, which means that the syringe and the push rod are relatively fragile. The moving distance of the push rod can be controlled in a plurality of modes, for example, the power assembly is subjected to preset moving distance, the moving distance of the push rod is indirectly controlled by controlling the power output of the power assembly, or an induction device is arranged on the moving path of the push rod so as to directly detect the moving distance of the push rod, and the mode can be more accurate and timely.

Above-mentioned induction system can adopt the form of sensor, for example photoelectric sensor is provided with the response target on the push rod, is provided with the response subassembly on the moving path of push rod, and when the descending critical position of push rod, the response subassembly can sense the response target on the push rod, and then produces sensing signal to learn that the push rod has arrived critical position. The critical positions of the push rod mainly comprise two, namely a descending limit, namely the position when the push rod pushes all reagents in the injector out, and the overlapping area of the push rod and the injector is at the maximum value; the second is the ascending limit, namely the position when the push rod fully sucks the reagent in the injector, and the overlapping area of the push rod and the injector is at the minimum value at the moment. Because the limit position of the push rod can be controlled in a plurality of simpler modes when the push rod moves upwards, if the limit position of the push rod moves upwards is provided with the stop block, the sensing device is generally arranged at the limit position of the push rod, and the push rod moves downwards to contact the bottom of the injector, so that the stop block and other structures are inconvenient to arrange.

Considering that the reagent holding chamber is preferably in the form of a syringe, which typically has only one reagent connection port through which both the addition of reagent to the syringe and the addition of reagent from the syringe to the chip are carried out, a reagent interface assembly may be provided at the reagent connection port of the syringe, through which the reagent connection port of the syringe may be brought into fluid communication with the reagent storage means and the chip, respectively. If the reagent interface assembly is provided with a main connecting channel, and a reagent input channel and a reagent output channel which are respectively communicated with the main connecting channel in a fluid manner, wherein the main connecting channel is communicated with the reagent connecting port on the injector in a fluid manner, the reagent input channel is communicated with the reagent storage device in a fluid manner, and the reagent output channel is communicated with the chip in a fluid manner, so that the connection of the injector, the reagent storage device and the chip can be realized.

When reagent is required to be added into the injector, the electromagnetic valve between the injector and the reagent storage device is opened, the electromagnetic valve between the injector and the chip is closed, the reagent storage device, the reagent interface assembly and the injector are in fluid communication, the push rod is driven by the power assembly to move upwards, and the reagent in the reagent storage device can be sucked into the injector through the reagent input channel on the reagent interface assembly; when reagent is needed to be filled into the chip, the electromagnetic valve between the injector and the chip is opened, the electromagnetic valve between the injector and the reagent storage device is closed, the injector, the reagent interface assembly and the chip are in fluid communication, the push rod is driven to move downwards through the power assembly, and the reagent in the injector can be pushed into the chip through the reagent output channel on the reagent interface assembly. Because the whole reagent sucking and pushing system is relatively closed, air can be effectively prevented from entering, and air bubbles generated in the injector are avoided.

Because a plurality of reactions are integrated on the microfluidic chip, the types of reagents required on the chip are also various, in order to realize the automatic filling of the various reagents on the chip, a plurality of groups of reagent accommodating chambers and reagent pushing devices can be arranged in an array manner, each group of reagent accommodating chambers and reagent pushing devices are relatively independent and are respectively communicated with different reagent storage devices and different reagent inlets on the chip in a fluid manner so as to realize the filling of various reagents, and each group of reagent accommodating chambers and reagent pushing devices are also independently controlled so as to realize the control of the filling of the various reagents. Of course, if each reagent is added sequentially, the situation that two or more reagents are added simultaneously does not occur, and all the reagent pushing devices and the reagent accommodating chambers can be controlled by one power assembly, so that the space can be saved and the complication of the structure can be avoided.

Separation detection system

The separation detection system comprises a capillary electrophoresis separation device and a fluorescence detection device, wherein the capillary electrophoresis separation device is gel electrophoresis generation equipment and is used for realizing the electrophoresis separation of DNA fragments; the fluorescence detection device is matched with the capillary electrophoresis separation device for use so as to realize the fluorescence detection of the DNA fragments.

In particular, a capillary electrophoresis separation system includes a capillary electrophoresis assembly, a temperature control assembly, a quick lock assembly, and a gel advancing device for effecting gel injection in a capillary. The capillary electrophoresis component is used for accommodating the capillary, provides a stable and reliable environment for electrophoresis separation, and avoids the damage of the capillary in the using process or in the replacement process; the temperature control assembly can provide temperature control for the capillary electrophoresis assembly during electrophoresis separation so as to ensure the smooth running of the electrophoresis separation; the quick locking assembly is used for temporarily locking the capillary electrophoresis assembly in the integral equipment so as to facilitate the replacement of the capillary.

The capillary electrophoresis device includes a capillary and a protection device for protecting the capillary, the capillary can be made of a commercially available product, such as an elastic fused silica capillary manufactured by Polymicro Technologies (PT), which is generally drawn from a semi-finished product of artificial fused silica, and similarly can be made of natural quartz, borosilicate glass, various plastic or fused silicon material, which mainly depends on the actual application requirements.

The capillary tube is preferably bent and arranged inside the protective component as a carrier for electrophoretic separation to form a smooth arc, so as to achieve the best effect of electrophoretic separation. In addition, generally, only one capillary is provided inside one capillary electrophoresis module, but sometimes, in order to improve the analysis throughput or the analysis efficiency, a plurality of capillaries are arranged in parallel in a capillary bundle.

The outside protection component of capillary mainly plays the effect of protection capillary, and its concrete form can adopt the protection casing, and the capillary is directly place in the inside of protection casing, and the protection casing and capillary form a whole, not only can play the effect that the damage was avoided to the protection capillary like this, but also can effectively improve the change mode of capillary. If bare capillaries are used directly in capillary electrophoresis separation systems, since capillaries are consumables, great care is taken to avoid breaking or crushing the capillaries when replacement is required, but when the capillaries are disposed inside a protective housing to form a unitary body, the unitary assembly can be replaced directly.

In the invention, the protection component is arranged on the outer side of the capillary tube, so that in order to meet the requirement of subsequent detection, a detection window is arranged on the protection component, and the fluorescence detection device can detect and analyze the electrophoretic separation in the capillary tube through the detection window. The fluorescence detection device of the invention is provided with a light path adjusting component besides traditional components such as a laser, an objective lens, a spectrometer and the like in the existing equipment, the light path adjusting component comprises a plane reflector, a lens or a filter lens and the like, an optical path between the laser and the objective lens can be changed through the light path adjusting component, and the miniaturization requirement is achieved through the relative position arrangement between the components.

The capillary electrophoresis component is replaceably arranged on the whole equipment, and in order to facilitate replacement, the whole equipment is provided with the quick locking component, the capillary electrophoresis component can be quickly locked on the whole equipment by utilizing the quick locking component, and when the capillary electrophoresis component needs to be replaced, the capillary electrophoresis component can be conveniently taken down from the whole equipment. The rapid locking assembly can be an existing elastic clamping and locking structure, the capillary electrophoresis assembly can be directly inserted into the elastic clamping and locking structure to realize locking, or the rapid locking assembly can also be a common thread locking structure, the capillary electrophoresis assembly can be directly screwed into the thread locking structure to realize locking, and of course, similar structures are numerous, so long as rapid locking can be realized.

Correspondingly, a structure matched with the quick locking assembly is also arranged on the capillary electrophoresis assembly, if the quick locking assembly adopts an elastic clamping and locking structure controlled by a solenoid assembly, a locking hole buckled with the locking head assembly needs to be arranged on the capillary electrophoresis assembly, the position of the locking hole is determined according to the installation position of the capillary electrophoresis assembly on the whole equipment, and the locking hole is generally arranged at the edge of the capillary electrophoresis assembly. Therefore, the locking structure matched with the capillary electrophoresis assembly can realize the quick replacement of the capillary electrophoresis assembly.

The capillary electrophoresis process has special requirements on temperature, for example, the temperature can influence separation reproducibility and separation efficiency, the control of the temperature can regulate the magnitude of electroosmotic flow, the temperature is increased, the viscosity of a buffer solution is reduced, the silicon light-based dissociation capability of a tube wall is enhanced, the electroosmosis speed is increased, the analysis time is shortened, the analysis efficiency is improved, but if the temperature is too high, the radial temperature difference in a capillary column is increased, the joule heating effect is enhanced, the column effect is reduced, and the separation efficiency is also reduced. Therefore, the capillary electrophoresis separation system is also provided with a temperature control assembly, and the temperature control assembly mainly adjusts the temperature of the capillary to ensure the smooth running of the electrophoresis process.

The capillary tube is arranged inside the protection assembly, so that the temperature control is not similar to the conventional mode, for example, a mode of directly sticking the temperature control assembly on the outer side of the protection assembly can be adopted, the mode not only can shorten a heat transfer path, and is convenient for quickly adjusting and controlling the temperature of the capillary tube, thereby greatly reducing the experiment time, but also having the effect of supporting the capillary tube electrophoresis assembly, and the temperature control assembly is generally directly fixed on the whole.

The temperature control assembly is preferably positioned on one side of the capillary electrophoresis assembly on the whole equipment, the shape of the temperature control assembly is matched with that of the capillary electrophoresis assembly, when the capillary electrophoresis assembly is locked on the whole equipment, the temperature control assembly is attached to the capillary electrophoresis assembly, and the temperature of the capillary is controlled by utilizing contact heat conduction. Wherein, the heating element in the temperature control assembly can adopt the common heating structure in this field, like resistance-type heating plate, and preferably adopts ultra-thin resistance-type heating plate to under the circumstances of realizing required heating effect, can show the reduction energy consumption and save space, provide support for the miniaturization of whole equipment.

Because whole equipment's miniaturized characteristics, the distance of each part can be less relatively in the equipment, in order to avoid causing the influence to whole equipment's other parts, heating element's outside should be provided with the isolation subassembly, and the isolation subassembly mainly can play the support heating element to and provide relatively confined environment for heating element. The arrangement mode of the isolation component can be made into an integral structure or a detachable structure and the like by analogy with the arrangement mode of the protective shell outside the capillary tube, and the isolation component of the temperature control component and the protective component of the capillary electrophoresis component can be contacted with each other and perform heat transfer when in use.

In order to ensure the heat transfer effect, the isolation component of the temperature control component and the protection component of the capillary electrophoresis component are made of materials with a relatively good heat transfer effect, and the capillary and the heating component are close to the side where the capillary and the heating component are contacted with each other as much as possible so as to shorten the heat transfer path. Meanwhile, a heat conduction gasket can be arranged between the temperature control assembly and the capillary electrophoresis assembly, the heat conduction gasket can be arranged on the capillary electrophoresis assembly and also can be arranged on the temperature control assembly, and the heat conduction gasket is preferably arranged on the capillary electrophoresis assembly so as to uniformly transfer the temperature to the capillary in consideration of the using effect.

In order to realize the automatic injection of the gel in the capillary tube, the gel propelling device is further provided in the invention, wherein the gel propelling device comprises a gel containing chamber which is used for temporarily storing the gel to be injected onto the capillary tube. Generally, in the field, the gel containing chamber is in the form of a syringe, so that the convenience in operation during gel filling is ensured, the structure is simplified as much as possible, and the filling amount of the gel can be well controlled.

The reagent inlet of the gel receiving chamber is in fluid communication with the gel storage device and the reagent outlet is in fluid communication with the capillary. The gel in the gel storage device can be added into the gel accommodating chamber in advance, and then the gel is timely filled into the capillary according to the requirement of the capillary electrophoresis system. The reagent accommodating chamber is provided with a temperature adjusting assembly, the temperature adjusting assembly can adjust the temperature of gel in the reagent accommodating chamber so as to enable the gel to be at an ideal storage temperature and facilitate temporary low-temperature storage of the gel, and the temperature adjusting assembly is preferably in the form of a semiconductor refrigerator based on the specific structure of the reagent accommodating chamber and the characteristics of the gel.

The temperature regulating assembly may be in direct contact with the reagent holding chamber to ensure its temperature control effect, e.g. when the reagent holding chamber is in the form of a syringe, the temperature regulating assembly may be disposed around the outside of the syringe to directly control the temperature of the gel in the syringe. However, in view of stability of the reagent-accommodating chamber during storage of the gel and injection, it is preferable that a support member is provided between the temperature regulating member and the reagent-accommodating chamber, such as a support member directly wrapped around the outside of the syringe, and the temperature regulating member is in contact with the support member, and heat is transferred to the reagent-accommodating chamber through the support member, which may slow down the temperature control speed, but is preferable in the present invention in view of stability of the whole apparatus.

In order that the temperature of the gel in the reagent holding chamber can be kept within a proper range for a long time, the outside of the reagent holding chamber may be provided with a temperature keeping assembly, which is preferably in direct contact with the reagent holding chamber, i.e., between the support assembly and the reagent holding chamber, such as the outside of the syringe, is directly wound around the temperature keeping assembly to keep the temperature of the gel in the syringe for a long time. Certainly, the heat preservation subassembly can also cause the influence to temperature regulation subassembly's heat transfer when maintaining the gel temperature in the reagent holds the chamber, but comparatively speaking, directly set up the whole result of use that the heat preservation subassembly can more be favorable to equipment in the outside that the chamber was held to the reagent, but also can play the effect that the chamber was held to the protection reagent, avoid supporting component etc. to hold the chamber and cause the damage to the reagent.

Because heat transfer's demand, reagent holds that chamber, thermal insulation component, supporting component and temperature regulation subassembly can be inseparable the setting together, should set up the protection component in its holistic outside to prevent that each subassembly from receiving external interference and damage. Wherein, the protection component can hold chamber, heat preservation component, supporting component and the whole cladding of temperature regulation subassembly and set up with reagent to guarantee the safeguard effect, from this, the chamber is held to reagent just needs to be connected through connecting tube and reagent storage device and capillary fluid, and this connecting tube also should satisfy heat retaining demand, takes place to change in order to prevent that the gel temperature from flowing through this connecting tube time, especially the connecting tube between chamber and the capillary is held to reagent. It should be noted that if the reagent accommodating chamber is in the form of a syringe, the heat preservation assembly, the support assembly, the temperature regulation assembly and the protection assembly can be only arranged on the main body part of the syringe, so that the problem that the overall use effect is influenced due to too complicated structure is avoided.

Since the reagent holding chamber will typically take the form of a syringe, when it is desired to add gel to the syringe or to fill the capillary with gel from the syringe, this can be done by pushing the plunger of the syringe, e.g. the upward movement of the plunger draws gel from the reagent storage device into the syringe and the downward movement of the plunger pushes gel from the syringe into the capillary. In view of the need for automation, the plunger of the injector may be coupled to a power assembly that automatically drives movement of the plunger in accordance with predetermined conditions. The power assembly generally adopts a combination form of a motor and a sliding block, the sliding block is fixedly connected with a push rod of the injector, and the motor can drive the sliding block to slide on a corresponding sliding rail, so that the push rod of the injector is driven to move upwards or downwards.

The integrated DNA analysis system integrates all steps of DNA detection and analysis on one device, is convenient to operate and rapid in detection, greatly improves the efficiency of DNA detection on the premise of ensuring the accuracy, can be widely applied to the fields of public security, justice, clinic and the like, and can fully meet the requirements of simplicity in operation and rapidness in detection.

A preferred embodiment of the integrated DNA analysis system of the present invention will be described with reference to FIGS. 1 to 36.

As shown in fig. 1 to 5, the integrated DNA analysis system of the present invention includes a reagent supply system, a microfluidic reaction system, and a separation detection system. The reagent supply system comprises a reagent storage device 100 and a reagent injection device 200, the microfluidic reaction system comprises a microfluidic chip 300 and a microfluidic device 400, the separation detection system comprises a capillary electrophoresis device 500 and a fluorescence detection device 600, specifically, the reagent storage device 100 is connected with the microfluidic chip 300 through the reagent injection device 200, a reagent in the reagent storage device 100 can be conveyed onto the microfluidic chip 300 through the reagent injection device 200 to complete corresponding operations, the flow of the reagent on the microfluidic chip 300 is controlled by the microfluidic device 400, a reactant on the microfluidic chip 300 can flow into the capillary electrophoresis device 500 to perform electrophoretic separation, and a result is detected through the fluorescence detection device 600 which is used in a matched mode.

As shown in fig. 6 to 15, the microfluidic chip 300 mainly integrates two steps of DNA extraction and PCR amplification, and includes a top plate 301, a fitting plate 302, a channel plate 303, a channel plate 304, a valve hole plate 305, a bottom plate 306, and a reaction plate 307, which are sequentially stacked. The top plate 301, the accessory plate 302, the pipeline plate 303, the channel plate 304, the valve hole plate 305 and the bottom plate 306 are identical in size and shape and are overlapped to form a chip body, the chip body is a bearing component of the microfluidic chip 300, and a sample extraction area, a fluid pipeline and a waste liquid discharge area are integrated on the chip body; the reaction plate 307 is attached to the chip body and is a reaction component of the microfluidic chip 300, a reaction region is formed on the reaction plate 307, and the reaction region on the reaction plate 307 is communicated with a fluid pipeline on the chip body.

Wherein, preferably, the top plate 301, the accessory piece 302, the pipeline piece 303, the channel piece 304, the valve hole piece 305, the material of film 306 that make up the chip body select the PET material (Polyethylene terephthalate), the PET material has good physical mechanical properties in the wider temperature range, long-term service temperature can reach 120 ℃, electrical insulation is good, even under high temperature high frequency, its electrical property is still better, creep resistance, fatigue resistance, friction resistance, dimensional stability are all fine, be particularly suitable for as the main material of chip, have the advantage of convenient processing, low price.

Meanwhile, it is preferable that the material of the reaction sheet 307 is PP (Polypropylene), wherein the PP has low density, strength, rigidity, hardness, and heat resistance superior to those of low-pressure polyethylene, can be used at about 150 ℃, has good dielectric properties and high-frequency insulation properties, is not affected by humidity, but becomes brittle at low temperature, is not wear-resistant, and is easy to age. Although PP is difficult to process and is not suitable for the main material of the chip, in the present embodiment, the reaction plate 307 is made of PP because the reaction efficiency can be significantly improved by using PP for the PCR reaction to make the reaction vessel.

Specifically, the top sheet 301 is an upper packaging sheet of the microfluidic chip 300, and mainly plays a role in sealing the chip body. In the present embodiment, a sample addition port 308, a hydrophobic air vent 309, a reaction plate heat dissipation port 310, a reagent addition port 311, and a reactant delivery port 312 are formed on the top plate 301 corresponding to the internal structure of the microfluidic chip 300.

The sample adding port 308 is communicated with a sample extraction area on the chip body, a blood sample to be processed can be added to the chip through the sample adding port 308, and a sealing cover plate 313 is arranged at the sample adding port 308; the hydrophobic exhaust port 309 is communicated with a hydrophobic exhaust area on a fluid pipeline in the chip body, and a hydrophobic exhaust fitting 314 is arranged at the hydrophobic exhaust port 309 for exhausting in the fluid pipeline; the reaction plate heat dissipation port 310 is communicated with the reaction plate 307 attached to the chip body, and is used for communicating the reaction plate 307 with the external environment so as to facilitate heat dissipation; the reagent adding ports 311 are communicated with reagent inlets on fluid pipelines in the chip body and are used for adding reaction reagents in the fluid pipelines, wherein the number of the reagent adding ports 311 is multiple, and each reagent adding port 311 corresponds to one reagent; the reactant delivery port 312 is provided with a connecting conduit 315, and the reactant delivery port 312 can be communicated with a subsequent reaction device (capillary) through the connecting conduit 315, so as to deliver the reactant on the microfluidic chip 300 to the subsequent device.

The fitting sheet 302 is located below the top sheet 301, and is used to provide mounting spaces for fittings on the microfluidic chip 300, such as a sample extraction fitting 316, a waste liquid discharge fitting 317, and the like. In consideration of the difficulty of processing, the thickness of the accessory piece 302 is not likely to be too large, so in order to ensure that each accessory has a sufficient installation space, the number of the accessory pieces 302 may be multiple, for example, two, three, etc., and two accessory pieces 302 are provided in this embodiment.

Corresponding to the internal structure of the microfluidic chip 300 and the structure of the top sheet 301, a sample extraction fitting mounting area 318, a hydrophobic air vent 309, a reaction plate heat dissipation opening 310, a reagent addition opening 311, a reactant delivery opening 312, and a waste liquid discharge fitting mounting area 319 are formed on the fitting sheet 302. Wherein the sample extraction fitting mounting area 318 corresponds to the sample addition port 308 on the top sheet 301 for receiving the sample extraction fitting 316; waste fitting mounting section 319 is for housing waste fitting 317.

It should be noted that in this embodiment, the sample collection assembly 316 is preferably an FTA strip, which is a proprietary technology of Whatman corporation, and is originally applied to DNA and RNA collection, transportation, purification and storage at room temperature, all on a single card. Of course, the sample extraction unit 316 may be a similar magnetic bead extraction unit, as long as it can extract a sample. In addition, the waste liquid discharge fitting 317 is preferably made of paper or the like having a fluid absorbing function for easy installation.

The duct chip 303 is located below the fitting chip 302 and is the carrier chip for the fluid channels 320, and the fluid agent flows mainly in the fluid channels 320 on the duct chip 303 to achieve flow between the functional regions. Meanwhile, the duct piece 303 also provides an installation space for each fitting together with the fitting piece 302 to facilitate connection of each fitting to the fluid pipe 320.

Corresponding to the internal structure of the microfluidic chip 300 and the structure of the fitting piece 302 described above in this embodiment, the channel piece 303 is formed with a fluid conduit 320, a sample extraction fitting mounting area 318, a reaction piece heat dissipation port 310, and a waste liquid discharge fitting mounting area 319. Wherein the sample extraction fitting mounting region 318 on the fitting piece 302 and the duct piece 303, and the sample extraction fitting 316 together form a sample extraction region; the waste fitting mounting section 319 on the fitting piece 302 and the duct piece 303 and the waste fitting 317 together form a waste discharge area; the fluid pipeline 320 is provided with a reagent inlet, a hydrophobic exhaust area and a reactant outlet, and the fluid pipeline 320 can connect the functional areas to facilitate the flow of the reaction fluid between the functional areas.

The reagent inlets of the fluid line 320 include an eluent inlet 321 and a PCR reagent inlet 322 corresponding to the DNA extraction and PCR amplification operations integrated on the microfluidic chip 300, and the reagent inlets of the fluid line 320 may further include a label inlet 323 corresponding to the subsequent electrophoresis and detection operations. Wherein, the eluent inlet 321 on the fluid pipeline 320 is communicated with the sample extraction region, so as to flush out the reactant extracted from the sample extraction region by the eluent input from the eluent inlet 321; the PCR reagent inlet 322 is communicated with the reaction area on the reaction plate 307, and the PCR reagent inputted from the PCR reagent inlet 322 can flow into the reaction area on the reaction plate 307 together with the reactant flushed from the sample extraction area to perform PCR reaction; the marker inlet 323 is connected to the reactant outlet 324 of the fluid line 320, and the marker can be fed from the marker inlet 323, then fed to the reactant outlet 324 along the fluid line 320, and fed to the subsequent reaction equipment through the reactant outlet 324, the reactant feeding port 312 and the connecting conduit 315.

In this embodiment, the fluid conduits 320 on the duct pieces 303 include: a first fluid conduit 325 connecting the sample extraction region and the hydrophobic vent region; a second fluid line 326 connecting the PCR reagent inlet 322 with the hydrophobic vent region; a third fluid line 327 connecting the hydrophobic vent region with the reaction region; a fourth fluid line 328 connecting the reaction zone with the reactant outlet 324; a fifth fluid conduit 329 connecting the label inlet 323 and the reactant outlet 324; and a waste liquid discharge line 330 connecting the sample extraction region and the waste liquid discharge region, the reaction region and the waste liquid discharge region, the reactant outlet 324 and the waste liquid discharge region. Of these, the third fluid line 327 connecting the hydrophobic vent region and the reaction region is preferably two to facilitate sufficient filling of the reaction region with reactants.

In this embodiment, the fluid channel 320 between any two functional regions on the channel plate 303 is not through but disconnected, that is, a channel disconnection point is formed on the fluid channel 320, and the fluid cannot directly flow between the functional regions through the fluid channel 320 on the channel plate 303, but the fluid flow on the microfluidic chip 300 is accurately controlled by the channel plate 304 and the valve hole 305 described below.

The channel plate 304 is located below the channel plate 303, and is used to provide a transition connection for the fluid pipeline 320 on the channel plate 303 to pass through, so as to precisely control the flow of the fluid on the microfluidic chip 300. Corresponding to the internal structure of the microfluidic chip 300 and the structure of the channel plate 303, a transitional connection hole set 331, a reaction region connection hole 332, and a reaction plate heat sink 310 are formed on the channel plate 304.

The transitional connection hole groups 331 are arranged corresponding to the pipeline disconnection points of the fluid pipelines 320 on the pipeline sheets 303, each transitional connection hole group 331 comprises two adjacent connection holes, and each connection hole is respectively communicated with the fluid pipelines 320 on two sides of the pipeline disconnection point on the fluid pipeline 320; the reaction zone connection holes 332 communicate with the reaction zones on the reaction plate 307 for delivering the reaction fluid to the reaction zones on the reaction plate 307.

The valve hole plate 305 is located below the channel plate 304 and is used for communicating the fluid pipeline 320 on the pipeline plate 303. Corresponding to the internal structure of the microfluidic chip 300 in this embodiment and the structures of the duct plate 303 and the channel plate 304, a valve hole 333, a reaction region connection hole 332, and a reaction plate heat dissipation port 310 are formed on the valve hole plate 305.

The valve holes 333 are arranged corresponding to the transitional connection hole groups 331 on the channel plate 304, each transitional connection hole group 331 is communicated with one valve hole 333, that is, two adjacent connection holes in the transitional connection hole group 331 are respectively communicated with the valve holes 333, so that the fluid pipeline 320 can be communicated at a pipeline disconnection point through the transitional connection hole group 331 and the valve holes 333.

Specifically, a first valve hole 334 is formed corresponding to the first fluid pipe 325 connecting the sample extraction area and the hydrophobic exhaust area; a second valve hole 335 is formed corresponding to the third fluid pipe 327 connecting the hydrophobic exhaust region and the reaction region; a third valve hole 336 is formed corresponding to the fourth fluid pipe 328 connecting the reaction region with the reactant outlet 324; a fourth valve hole 337 is formed corresponding to the waste liquid discharge line 330 connecting the sample taking area and the waste liquid discharge area, the reaction area and the waste liquid discharge area, and the reactant outlet 324 and the waste liquid discharge area.

The bottom plate 306 is located below the valve hole plate 305 and is a lower packaging plate of the microfluidic chip 300. Corresponding to the internal structure of the microfluidic chip 300 and the structure of the valve hole plate 305, the reaction region connecting hole 332 and the reaction plate heat dissipation hole 310 are formed on the bottom plate 306.

Meanwhile, in order to control the opening and closing of the valve hole 333 in the valve hole piece 305, the base plate 306 preferably has elasticity, so that the base plate 306 can be deformed by applying an external force to a position of the base plate 306 corresponding to the valve hole 333 in the valve hole piece 305, and the base plate 306 can block the valve hole 333 in the valve hole piece 305, so that the fluid pipeline 320 is cut off at the pipeline cutting point. It should be noted that the external force applied to the bottom plate 306 can be applied by the microfluidic device 400 used with the microfluidic chip 300, for example, a solenoid valve is disposed in the microfluidic device 400 corresponding to the valve hole 333 on the valve hole 305, and the opening and closing of the valve hole 333 can be realized by the lifting and lowering of the solenoid valve.

The reaction sheet 307 is located below the bottom sheet 306, and is a special reaction container, the size and shape of which can be flexibly set according to different situations, and in addition, a protective gasket 338 can be arranged between the reaction sheet 307 and the bottom sheet 306. Corresponding to the internal structure of the microfluidic chip 300 and the structure of the chip body, a reaction region 339, a reactant inlet 340 and a reactant outlet 341 are formed on the reaction sheet 307.

Among them, in order to enhance the effect of the PCR reaction, the shape of the reaction region 339 on the reaction piece 307 is preferably S-shaped. The S-shaped reaction region 339 is communicated with the corresponding reaction region 339 connecting hole 332 on the chip body through the reactant inlet 340, so that the reactant can flow into the reaction region 339 through the fluid conduit 320, the reaction region connecting hole 332 and the reactant inlet 340 to perform a PCR reaction. The reacted product can be delivered to the reactant outlet 324 on the channel plate through the reactant outlet 341, the corresponding reaction region connecting hole 332 on the chip body, and the fluid pipeline 320, and then delivered to the subsequent reaction equipment through the reactant delivery port 312 and the connecting conduit 315.

The working process of the microfluidic chip 300 of the present invention is: first, an external force is applied to the corresponding region of the bottom plate 306 to close all the valve holes 333 of the valve hole plate 305, so that the corresponding fluid conduits 320 of the channel plate 304 are in a disconnected state; secondly, the blood sample to be tested is added to the sample extraction fitting 316 in the sample extraction area through the sample adding port 308 on the top sheet 301; then, the first valve hole 334 of the valve hole piece 305 is opened to make the first fluid pipeline 325 between the sample extraction area and the hydrophobic exhaust area in a through state, and simultaneously, the eluent is introduced from the eluent inlet 321, so that the DNA can be eluted from the sample extraction fitting 316 in the sample extraction area and flow to the hydrophobic exhaust area; then, the first valve hole 334 of the valve hole piece 305 is closed, the second valve hole 335 is opened, the third fluid conduit 327 between the hydrophobic exhaust region and the reaction region 339 is in a through state, and the DNA and the PCR reagent enter the reaction region 339 on the reaction piece 307 through the third fluid conduit 327; then, the second valve hole 335 on the hole plate 305 is closed to make the reaction area 339 on the reaction plate 307 be an isolated chamber for performing PCR reaction; finally, after the PCR reaction is completed, the third valve hole 336 is opened to allow the reaction region 339 to be in a communicating state with the fourth fluid conduit 328 of the reactant outlet 324, and the reactant in the reaction region 339 can flow to the reactant outlet 324 while the label is introduced from the label inlet 323, so that the amplified DNA and the label can enter the subsequent capillary for electrophoretic separation. In the whole process, if the waste liquid is excessive, the fourth valve hole 337 can be selectively opened, so that the waste liquid discharge pipe 330 between the sample extraction region and the waste liquid discharge region, between the reaction region 339 and the waste liquid discharge region, and between the reactant outlet 324 and the waste liquid discharge region can be selectively communicated, and the redundant fluid can enter the waste liquid discharge region.

As shown in fig. 16 to 18, the microfluidic device 400 includes a microfluidic control box 401 and a mechanical control platform 402, wherein the microfluidic control box 401 can carry the microfluidic chip 300, control the flow of fluid in the chip, and control the reaction temperature on the chip; the mechanical control platform 402 may enable movement of the microfluidic control cartridge 401, thereby making chip replacement easier.

Further, the microfluidic control cartridge 401 includes a chip platform 403 and a carrying bottom frame 404, wherein the carrying bottom frame 404 constitutes a main body frame of the microfluidic control cartridge 401, and the chip platform 403 is disposed on the carrying bottom frame 404 for carrying a chip. The chip platform 403 is a plate structure, a chip accommodating groove 405 is formed in the top surface of the plate structure, the chip accommodating groove 405 is an open structure, the shape of the chip accommodating groove 405 is matched with that of the chip, and the chip can be directly placed in the chip accommodating groove 405. Preferably, since the chip is loaded at a position in the chip accommodating recess 405 with high accuracy to ensure the subsequent flow control of the fluid, a position calibration assembly, such as a cylindrical protrusion corresponding to the recess on the chip, may be further disposed in the chip accommodating recess 405, so that the chip can be placed more accurately and easily by using the position calibration assembly.

A chip locking device 406 is disposed at one side of the chip receiving recess 405, the chip locking device 406 is of a conventional rotary locking structure, and after the chip is placed in the chip receiving recess 405, the chip locking device 406 can be rotated so that the chip is firmly pressed in the chip receiving recess 405 by the chip locking device 406, thereby ensuring the chip loading firmness and the chip stability during operation. Optionally, an elastic locking structure may be directly disposed in the chip accommodating recess 405, and the chip may be locked after being placed in the chip accommodating recess 405, so that the chip may be more conveniently replaced, and the chip platform 403 may be prevented from being provided with redundant components, thereby saving space.

The chip accommodating recess 405 is provided with a temperature control component 407, wherein the temperature control component 407 is disposed on the bottom surface of the chip accommodating recess 405 and corresponds to the reaction plate 307 on the microfluidic chip 300, and after the chip is placed in the chip accommodating recess 405, the temperature control component 407 directly contacts the reaction plate 307 on the microfluidic chip 300 to adjust the temperature of the reaction region 339 on the reaction plate 307, thereby ensuring the normal reaction. In this embodiment, the temperature control assembly 407 includes a thermal pad, a semiconductor cooler, and a temperature sensor disposed between the thermal pad and the semiconductor cooler, the thermal pad directly contacts with the chip after the chip is placed in the chip receiving recess 405, and the semiconductor cooler adjusts the reaction temperature on the chip.

A plurality of intermediate connecting channels 408 are further disposed on the bottom surface of the chip accommodating recess 405, the intermediate connecting channels 408 are disposed through the chip platform 403, wherein the plurality of intermediate connecting channels 408 on the bottom surface of the chip accommodating recess 405 are in one-to-one correspondence with the plurality of valve holes 333 on the chip. Correspondingly, a plurality of solenoid valves 409 are arranged on the bearing bottom frame 404 below the chip platform 403, wherein a plurality of solenoid valve mounting holes 410 are arranged on a bottom surface 414 of the bearing bottom frame 404, the solenoid valves 409 are vertically arranged in the solenoid valve mounting holes 410, the plurality of solenoid valves 409 are arranged to correspond to the plurality of intermediate connecting channels 408 on the bottom surface of the chip accommodating groove 405 one by one, and top plungers of the solenoid valves 409 pass through the corresponding intermediate connecting channels 408 and then contact the bottom plate 306 on the microfluidic chip 300, and can drive positions of corresponding valve holes 333 on the bottom plate 306 to lift up and down, so as to open or close the corresponding valve holes 333 on the chip.

The microfluidic control cartridge 401 is further provided with a heat sink 413, and the heat sink 413 is used for dissipating heat of the entire microfluidic control cartridge 401 to prevent it from failing to work due to high temperature. The heat dissipation device 413 comprises a mounting shell 412 and a heat dissipation fan 411 embedded on the mounting shell, the mounting shell 412 is arranged outside the bearing bottom frame 404 in a covering mode to form a relatively closed space, the solenoid valve 409 is located in the closed space, and the heat dissipation fan 411 is arranged on the side wall of the mounting shell 412, enables heat dissipation airflow to cover the solenoid valve 409, and is preferably arranged in an opposite mode to save space and improve heat dissipation efficiency.

The mechanical control platform 402 comprises a horizontal displacement device and a vertical displacement device, wherein the microfluidic control box 401 is fixedly arranged in the horizontal displacement device and the vertical displacement device, the movement of the microfluidic control box 401 in the horizontal direction can be realized through the horizontal displacement device, the movement of the microfluidic control box 401 in the vertical direction can be realized through the vertical displacement device,

the horizontal displacement device comprises a horizontal sliding part 415 and a first motor 416, the horizontal sliding part 415 comprises a sliding rail which is fixedly arranged and a sliding part which is matched with the sliding rail for use, the sliding rail can be selectively fixed on a base part and is used as a moving base, the sliding part is in sliding fit with the sliding rail, the first motor 416 is connected with the sliding part, and a bearing bottom frame 404 on the microfluidic control box 401 is fixedly arranged on the sliding part, so that the sliding part can drive the whole microfluidic control box 401 to reciprocate along the sliding rail under the driving of the first motor 416, and the horizontal movement of the microfluidic control box 401 is realized.

The vertical displacement device also comprises a vertical sliding part 417 and a second motor 418, the vertical sliding part 417 comprises a sliding rail fixedly arranged and a sliding part matched with the sliding rail for use, the sliding rail can be selectively fixed on a base part and is used as a moving base, the sliding part is in sliding fit with the sliding rail, the second motor 418 is connected with the sliding part, the microfluidic control box 401 and the horizontal displacement device are fixedly arranged on the sliding part together, and therefore, under the driving of the second motor 418, the sliding part can drive the whole microfluidic control box 4011 and the horizontal displacement device to reciprocate along the sliding rail, so that the vertical movement of the microfluidic control box 401 is realized.

In this embodiment, the vertical displacement device is required to drive the microfluidic control cartridge 401 and the horizontal displacement device to move together, so that the slide rail in the horizontal displacement device is fixedly arranged in the first frame 419, and the horizontal displacement device moves the microfluidic control cartridge 401 horizontally relative to the first frame 419. Meanwhile, the sliding part in the vertical displacement device is fixedly connected with the first frame 419, and the sliding rail is fixedly connected with the second frame 420, so that under the driving of the second motor 418, the sliding part in the vertical displacement device can drive the first frame 419, and the microfluidic control box 401 and the horizontal displacement device which are arranged on the first frame 419 to vertically move together relative to the second frame 420.

As shown in fig. 19 to 21, the reagent storage device 100 of the present invention includes two kinds, one is a conventional reagent cartridge 101 and one is a temperature-controlled reagent cartridge 102. The conventional reagent kit 101 is shown in fig. 19, and mainly includes a reagent bottle and a conduit for storing reagents that do not have too high a temperature requirement, such as buffers and the like; the temperature control kit 102 is shown in fig. 20 and 21, and includes a temperature-keeping box, a reagent bottle and a conduit for storing a reagent with a relatively high requirement on temperature, such as DNA polymerase.

Specifically, the heat preservation box body comprises a heat preservation sheath 103, a temperature control assembly 104 and a protection box body 105, wherein a reagent bottle 106 is fixed in the heat preservation sheath 103 and used for preserving liquid; the temperature control component 104 is fixed on the heat-insulating sheath 103 and is used for controlling the temperature of the liquid in the reagent bottle 106; an insulating sheath 103 is mounted within the protective casing 105 for insulating the reagent bottles 106.

The heat-insulating sheath 103 is formed by buckling two shells with semi-arc inner walls, the two shells are used for wrapping the bottle body of the reagent bottle 106, and the bottle mouth part of the reagent bottle 106 protrudes out of the upper end face of the heat-insulating sheath 103; the outer wall of heat preservation sheath 103 is provided with temperature control assembly 104, and this temperature control assembly 104 includes semiconductor refrigerator and temperature sensor, and semiconductor refrigerator has temperature adjustable, small, energy consumption advantage such as low.

The heat-insulating sheath 103 and the temperature control assembly 104 are arranged in the protective box body 105, the protective box body 105 comprises a front box body 107, a rear plate 108 and a box cover 109, the rear side of the front box body 107 is open, a gap is arranged at the top of the front box body 107, the heat-insulating sheath 103 is embedded in an inner cavity of the front box body 107, the gap is used for a bottle mouth part of the reagent bottle 106 to pass through, the rear plate 108 is fixed at the rear side of the front box body 107 to seal the open side of the front box body 107, the upper end of the rear plate 108 extends upwards, the box cover 109 is arranged at the gap of the front box body 107, the box cover 109 is used for sealing the bottle mouth part, the rear side of the box cover 109 is abutted to the extending part of the rear plate 108, and a guide pipe 111 arranged in the reagent bottle 106 extends out of the box cover 109. In addition, it is preferable that a backing plate 110 is provided between the rear plate 108 and the front box 107, a concave portion is provided on the backing plate 110, and the temperature control device is provided on the rear outer wall of the thermal insulation sheath 103, and the concave portion is used for accommodating the temperature control device.

As shown in fig. 22 to 25, the reagent injection device 200 includes a mounting panel 201, a plurality of syringes 202 are vertically arranged side by side on a first side of the mounting panel 201, each syringe 202 is connected with a reagent push rod 203, and the reagent push rod 203 can reciprocate inside the syringe 202 to suck a reagent into the syringe 202 or push the reagent inside the syringe 202 out. Wherein, each group of the injector 202 and the reagent push rod 203 are relatively independent, and a plurality of groups of the injectors 202 and the reagent push rods 203 which are arranged side by side can realize the filling of a plurality of reagents on the chip.

A reagent interface 204 is disposed on a lower portion of the first side of the mounting panel 201, and a set of a syringe connection channel 205, a reagent input channel 206 and a reagent output channel 207 is disposed on the reagent interface 204 corresponding to each syringe 202, wherein the syringe connection channel 205 is disposed on a top surface of the reagent interface 204, the reagent input channel 206 is disposed on a side surface of the reagent interface 204, the reagent output channel 207 is disposed on a bottom surface of the reagent interface 204, and the reagent input channel 206 and the reagent output channel 207 are respectively in fluid communication with the syringe connection channels 205.

Each syringe connection channel 205 on the reagent interface 204 corresponds to an upper syringe 202, the bottom of the syringe 202 is inserted into the syringe connection channel 205 on the reagent interface 204, and the reagent interface of the syringe 202 is in fluid communication with the syringe connection channel 205. The reagent input channel 206 on the reagent interface device 204 is in fluid communication with the reagent outlet on the reagent storage device 100, and a first solenoid valve 208 is disposed on the connection channel between the reagent input channel 206 and the reagent outlet on the reagent storage device 100; the reagent output channel 207 on the reagent interface device 204 is in fluid communication with the reagent inlet on the chip, and a second solenoid valve is also provided on the connection channel between the reagent output channel 207 and the reagent inlet on the chip.

Thus, the reagent interface on the syringe 202 is in fluid communication with the reagent outlet on the reagent storage device 100 through the syringe connection channel 205, the reagent input channel 206 on the reagent interface device 204; the reagent interface on the injector 202 is in fluid communication with the reagent inlet on the microfluidic chip 300 via the injector connecting channel 205 and the reagent output channel 207 on the reagent interface device 204. When the first solenoid valve 208 on the connection channel between the reagent input channel 206 and the reagent outlet on the reagent storage device 100 is opened, the reagent on the reagent storage device 100 can flow into the syringe 202 through the reagent outlet on the reagent storage device 100, the reagent input channel 206 on the reagent interface device 204, the syringe connection channel 205, and the reagent interface on the syringe 202, so as to realize the addition of the reagent in the syringe 202. When the second electromagnetic valve on the connection channel between the reagent output channel 207 and the reagent inlet on the chip is opened, the reagent in the injector 202 can flow into the chip through the reagent interface on the injector 202, the injector connection channel 205 on the reagent interface device 204, the reagent output channel 207 and the reagent inlet on the chip, so as to fill the reagent in the chip.

The bottom of the syringe 202 is inserted into the syringe connection channel 205 of the reagent interface 204, and not only the purpose of connecting the reagent storage apparatus 100 and the chip is achieved, but also the bottom of the syringe 202 is relatively fixedly disposed on the mounting panel 201. Meanwhile, the installation panel 201 is further provided with an injector clamping device 209 above the reagent interface device 204, a clamping hole is provided on the injector clamping device 209 at a position corresponding to each injector 202, the injector 202 can be clamped in the clamping hole on the injector clamping device 209, and the injector 202 can be firmly arranged on the installation panel 201 through the reagent interface device 204 and the injector clamping device 209. Preferably, the position of the clamping hole on the syringe clamping device 209 corresponds to the upper or middle upper portion of the syringe 202, such that the bottom of the syringe 202 is fixed by the reagent interface device 204 and the middle upper portion of the syringe 202 is fixed by the syringe clamping device 209, such that the stability of the syringe 202 is ensured when the reagent pusher 203 reciprocates in the syringe 202.

The reciprocating motion of the reagent pushing rod 203 in the injector 202 is realized by a motor 210, a sliding component 211 and a sliding track 212, wherein the motor 210 is connected with the sliding component 211, the sliding component 211 is slidably disposed on the sliding track 212, and the sliding component 211 is connected with the reagent pushing rod 203, so that the motor 210 can drive the sliding component 211 to reciprocate on the sliding track 212, and the sliding component 211 can drive the reagent pushing rod 203 to reciprocate in the injector 202. It should be noted that each set of syringes 202 and reagent push rods 203 is provided with a corresponding set of motors 210, slide assemblies 211, and slides 212 to facilitate individual control of each set of syringes 202 and reagent push rods 203.

In order to detect the limit position of the reagent pusher 203 during the reciprocating movement in the injector 202, an optical sensor 213 is disposed on the first side surface of the mounting panel 201, a detection piece 214 is disposed on the sliding assembly 211 corresponding to the optical sensor 213, and the optical sensor 213 and the detection piece 214 operate on the principle of photoelectric effect. The optical sensor 213 is disposed on a moving path of the reagent push rod 203, and corresponds to a downward limit position of the reagent push rod 203, when the sliding component 211 drives the reagent push rod 203 to move downward, the detection sheet 214 passes through the optical sensor 213 when the reagent push rod 203 reaches the downward limit position, and then generates a sensing signal in the optical sensor 213 to indicate that the reagent push rod 203 has reached the downward limit position, and at this time, the motor 210 stops driving.

The specific working process of the reagent injection device 200 is as follows: first, the first electromagnetic valve 208 on the connection channel between the reagent input channel 206 and the reagent outlet on the reagent storage device 100 is opened, and the second electromagnetic valve on the connection channel between the reagent output channel 207 and the reagent inlet on the chip is closed; secondly, the motor 210 drives the sliding assembly 211 to move upwards along the sliding rail 212, the sliding assembly 211 drives the reagent pushing rod 203 to move upwards in the injector 202, so that the reagent in the reagent storage device 100 is sucked into the injector 202, and after the reagent is added, the first electromagnetic valve 208 on the connecting channel between the reagent input channel 206 and the reagent outlet on the reagent storage device 100 is closed; then, according to the type of the reagent to be filled on the chip, opening a second electromagnetic valve on a connecting channel between the corresponding reagent output channel 207 and a reagent inlet on the chip; then, the motor 210 drives the sliding assembly 211 to move downward along the sliding rail 212, and the sliding assembly 211 drives the reagent pushing rod 203 to move downward inside the syringe 202, so as to push the reagent in the syringe 202 to the corresponding reagent inlet on the chip, thereby meeting the requirements of reaction or operation.

As shown in fig. 26 to 35, a capillary electrophoresis device 500 includes a capillary electrophoresis cassette 501, a capillary heating cassette 502, a quick locking mechanism 503, and a gel advancing device 504, wherein the capillary electrophoresis cassette 501 is an assembly accommodating a capillary 505, which is an execution part of electrophoresis separation; capillary heat box 502 is an assembly that houses a heating plate 506, a means for providing temperature regulation for electrophoretic separations; the quick locking mechanism 503 is a component for detachably locking the capillary electrophoresis cartridge 501 to the whole apparatus, so as to facilitate replacement of the capillary electrophoresis cartridge 501; gel advancing device 504 is used for automated injection of gel in capillary 505.

Further, the capillary electrophoresis cartridge 501 includes an electrophoresis cartridge body 507, the electrophoresis cartridge body 507 is sheet-shaped, an accommodation space is formed inside, the capillary tube 505 is disposed in the accommodation space of the electrophoresis cartridge body 507, and the electrophoresis cartridge body 507 can protect the capillary tube 505. The electrophoresis cartridge 507 is formed by combining a first housing 508 and a second housing 509, the capillary 505 is located between the first housing 508 and the second housing 509, and a protective gasket 510 may be further disposed between the first housing 508 and the capillary 505, and the protective gasket 510 may be a foam gasket or a gasket made of other elastic materials, so as to provide further protection for the capillary 505.

The capillary tube 505 is bent in a U-shape and the main part of the capillary tube 505 is positioned in the electrophoresis cartridge body 507 with the fluid inlet end of the capillary tube 505 having a length greater than the fluid outlet end, whereby the fluid inlet end of the capillary tube 505 can protrude from the electrophoresis cartridge body 507 when the capillary tube 505 is positioned in the electrophoresis cartridge body 507 to facilitate the filling of fluid in the capillary tube 505. Meanwhile, since the fluid inlet end of the capillary tube 505 protrudes from the electrophoresis cartridge body 507, a protective sleeve 511 is preferably sleeved on the fluid inlet end of the capillary tube 505, and the protective sleeve 511 can protect the fluid inlet end of the capillary tube 505 to prevent damage to the capillary tube 505 during use or movement.

The position corresponding to the fluid outlet end of the capillary 505 is provided with a detection window 512 on the electrophoresis cartridge 507, where the detection window 512 is for facilitating the observation of the electrophoresis separation in the capillary 505 by the fluorescence detection apparatus 600 in the capillary 505 electrophoresis apparatus 500, that is, the laser can enter through the detection window 512 for signal detection. Correspondingly, since the capillary tube 505 is relatively exposed at the detection window 512, the detection window 512 should be provided with a closing component 549, when detection is needed, the closing component 549 is removed to open the detection window 512, so that laser in the fluorescence detection device 600 can enter for signal detection, and after detection is finished, the closing component 549 is installed at the detection window 512 to close the detection window 512, so as to prevent the capillary tube 505 from being accidentally damaged.

One side that is close to capillary heating box 502 on the second casing 509 is provided with heat conduction gasket 513, and when capillary electrophoresis box 501 installed whole equipment, one side that is provided with heat conduction gasket 513 on capillary electrophoresis box 501 can laminate each other with capillary heating box 502, through setting up heat conduction gasket 513, can make capillary 505 be heated evenly in the course of the work, and the distance between heat conduction gasket 513 and the capillary 505 is shorter moreover, can improve heating efficiency greatly, and then improve experimental efficiency. In addition, the electrophoresis box body 507 can be further provided with a quick clamping groove 514, and the quick clamping groove 514 is used for being connected with a quick clamping piece on the whole equipment, so that the capillary electrophoresis box 501 can be quickly detached and installed on the whole equipment.

The capillary tube heating cassette 502 includes a heating cassette body 515, the shape of the heating cassette body 515 is preferably matched with the shape of the electrophoresis cassette body 507, and is also sheet-shaped, the inside of the heating cassette body 515 forms a housing chamber, the heating plate 506 is disposed in the housing chamber, the heating cassette body 515 can protect the heating plate 506, the heating plate 506 is prevented from being damaged, and the temperature of the heating plate 506 can be prevented from affecting other components.

Wherein the heating cassette body 515 includes a bottom plate 516, the bottom plate 516 has a heat transfer property, and when the heating cassette body 515 is in contact with the electrophoresis cassette body 507, the bottom plate 516 may be in contact with the heat conductive pad 513 on the electrophoresis cassette body 507 to transfer heat. Preferably, the heating plate 506 is fixedly disposed on the bottom plate 516 to minimize a heat transfer path with the capillary tube 505, the heating plate 506 is disposed at a middle position of the bottom plate 516 and is connected to a temperature sensor 517, and the temperature sensor 517 is connected to a control system through a signal connection 518, so as to realize real-time monitoring of the temperature of the heating plate 506. In the present embodiment, the heating plate 506 is preferably an ultra-thin resistance heating plate, and the thickness of the ultra-thin resistance heating plate can be designed to be 10-20mm, so that not only can the energy consumption be significantly reduced, but also the miniaturization can be achieved under the condition of achieving the required heating effect.

The bottom plate 516 is connected with a support frame 519, the support frame 519 is a frame-type structure, locking components 520 are correspondingly arranged at the edges of the bottom plate 516 and the support frame 519, and the bottom plate 516 and the support frame 519 can be fixedly connected through the locking components 520. A cavity is formed on the support frame 519 at a position corresponding to the heating plate 506, after the support frame 519 is connected with the bottom plate 516, the heating plate 506 on the bottom plate 516 is located at the middle cavity of the support frame 519, preferably, a protective cushion 521 is arranged between the bottom plate 516 and the support frame 519, the protective cushion 521 may be made of a foam board or other elastic materials, and the protective cushion 521 can protect the heating plate 506 and the temperature sensor 517.

After the bottom plate 516 and the support frame 519 are fixedly connected, a protective casing 522 can be sleeved on the outer side, the protective casing 522 can be connected with the bottom plate 516 and the support frame 519 in a buckling or clamping manner, and the protective casing 522 enables the heating plate 506 and the temperature sensor 517 on the bottom plate 516 to be in a relatively closed environment so as to avoid mutual interference with the external environment. In addition, a locking connector 523 may be further provided at an edge of the support frame 519, and the capillary heating cartridge 502 may be fixed to the overall apparatus by the locking connector 523.

The quick locking mechanism 503 includes a locking bottom frame 524, one end of the locking bottom frame 524 is provided with a locking swing groove 526, a locking member 525 is movably disposed in the locking swing groove 526, the locking member 525 is used in cooperation with the quick clamping groove 514 on the capillary electrophoresis cassette 501, and the locking member 525 can reciprocate in the locking swing groove 526 to release and lock the capillary electrophoresis cassette 501. Wherein, the bottom of the locking element 525 and the corresponding position on the locking swing groove 526 are respectively formed with a rotating shaft hole, a rotating shaft is arranged in the rotating shaft hole, and the locking element 525 can swing around the rotating shaft in a reciprocating way.

The other end of the locking bottom frame 524 is provided with a driving member mounting plate 527, and the end of the solenoid assembly 528 can pass through the mounting hole of the driving member mounting plate 527 and then be fixedly connected with the locking assembly 529, so as to fix the solenoid assembly 528 on the locking bottom frame 524. The end of the plunger 530 of the solenoid assembly 528 is provided with a driving connecting piece 531, the middle upper part of the latch 525 is provided with a driving connecting hole, the driving connecting piece 531 can be connected with the driving connecting hole on the latch 525, and the plunger 530 of the solenoid assembly 528 can drive the latch 525 to swing through the driving connecting piece 531 so as to release the capillary electrophoresis cassette 501.

A stop washer 532 is disposed on the plunger 530 of the solenoid assembly 528 at a position close to the locking member 525, one side of the stop washer 532 contacts the locking member 525, a return spring 533 is disposed between the other side and the driving member mounting plate 527, and the return spring 533 is sleeved on the plunger 530 of the solenoid assembly 528. Therefore, when the latch 525 needs to be opened, the plunger 530 of the solenoid assembly 528 can drive the latch 525 to swing through the driving connection member 531, and the latch 525 can contact the stop washer 532 and press the stop washer 532 and the return spring 533, so that the return spring 533 is in a compressed and deformed state; when the locking element 525 needs to be closed, the deformation force of the restoring spring 533 can be directly utilized to push the stop washer 532, so as to restore the locking element 525.

The gel advancing device 504 comprises a syringe 534, a protective cartridge 535, a semiconductor refrigerator 536 and an advancing assembly 537, wherein the syringe 534 is used to suck and advance the gel in the gel storage device into the capillary 505; the protective box 535 is arranged outside the syringe 534 and can protect the syringe 534; a semiconductor refrigerator 536 is provided inside the protective case 535, and the temperature of the gel in the syringe 534 can be adjusted; the pushing assembly 537 is connected to the syringe 534, and the syringe 534 can be automatically controlled to draw and push the gel in the reagent storage device 100 into the capillary 505.

The injector 534 includes a reagent cylinder 537 and a reagent push rod 538, wherein the reagent push rod 538 is inserted into the reagent cylinder 537 and reciprocally moves in the reagent cylinder 537, the reagent cylinder 537 is used to store gel, and the reagent push rod 538 is used to suck or push the gel into or out of the reagent cylinder 537.

The protection box 535 comprises a protection shell, a supporting block and a heat-insulating sheath 539, wherein the protection shell, the supporting block and the heat-insulating sheath 539 are sequentially arranged on the outer side of the reagent cylinder 537 in the injector 534, and the functions of adjusting the temperature of the gel in the reagent cylinder 537 and protecting the reagent cylinder 537 are achieved. Specifically, the heat-insulating sheath 539 is disposed outside the reagent cylinder 537, so that the gel in the reagent cylinder 537 can be kept at a proper temperature for a long time, and meanwhile, the reagent cylinder 537 can be prevented from being damaged by other components, such as being scratched or cracked by the supporting block. Wherein, the sheath 539 that keeps warm can be two arc parts, and the laminating of two curved sheaths 539 that keep warm sets up in the outside of reagent cylinder 537, and of course, the sheath 539 that keeps warm also can adopt the banded structure, and direct winding sets up in the outside of reagent cylinder 537.

The supporting block includes first supporting block 541 and second supporting block 542, and first supporting block 541 and second supporting block 542 detain and establish in the outside of reagent barrel 537 and heat preservation sheath 539, can play the tight effect of support clamp to reagent barrel 537, still can be with the inseparable laminating in the outside of reagent barrel 537 of heat preservation sheath 539 simultaneously. Of course, the first supporting block 541 and the second supporting block 542 may be made into an integral structure, and directly sleeved outside the reagent cylinder 537 and the thermal insulation sheath 539, but in consideration of convenience of assembly and disassembly, a split structure is preferred.

The second supporting block 542 is in contact with the semiconductor refrigerator 536, and heat generated from the semiconductor refrigerator 536 can be transferred to the reagent cylinder 537 through the second supporting block 542 to adjust the temperature of the gel in the reagent cylinder 537. Thus, the second supporting block 542 should have a heat conductive property, and a heat conductive pad 543 may be disposed between the second supporting block 542 and the semiconductor cooler 536 in consideration of uniformity of heat transfer. The semiconductor cooler 536 has the characteristics of small volume, low energy consumption and high temperature change rate, and the heat conducting gasket 543 is arranged between the semiconductor cooler 536 and the second supporting block 542, so that the heat conduction is more uniform, and the accuracy of controlling the chip reaction temperature is improved; meanwhile, the semiconductor refrigerator 536 may also be connected to a temperature sensor 517, and real-time temperature detection of the semiconductor refrigerator 536 may be achieved through the temperature sensor 517.

The protective casing comprises a first casing 544 and a second casing 545, and the first casing 544 and the second casing 545 are buckled on the outer sides of the support block and the semiconductor refrigerator 536, so that the support block, the semiconductor refrigerator 536, the insulating sheath 539 and the reagent cylinder 537 are in a relatively closed space, and are prevented from being interfered and damaged by the outside. The protective shell is designed into a split structure, and is mainly convenient to assemble and disassemble.

It should be noted that, in this embodiment, the protection housing is disposed outside the main body of the reagent cylinder 537, only the reagent interface and the push rod inlet on the reagent cylinder 537 are located outside the protection housing, the reagent interface on the reagent cylinder 537 is directly connected to the connection elbow 540, the flow direction of the gel can be changed through the connection elbow 540, and various pipe joints can be connected. The above-mentioned manner is mainly for convenience of connection and operation, and of course, the reagent cylinder 537 may be disposed entirely inside the protective casing, so that the reagent push rod 538 needs to pass through the protective casing and then be inserted into the reagent cylinder 537, and the connection elbow also needs to pass through the protective casing and then be connected to the reagent interface of the reagent cylinder 537.

The propelling assembly 537 comprises a slide rail 546, a slide 547 and a driving motor 548, wherein the driving motor 548 is connected with the slide 547, the slide 547 is slidably disposed on the slide rail 546, and the slide 547 is connected with the reagent push rod 538, so that the driving motor 548 can drive the slide 547 to reciprocate on the slide rail 546, and the slide 547 can drive the reagent push rod 538 to reciprocate on the reagent cylinder 537. Specifically, the slide rail 546 can be fixedly disposed on the protective case 535, and the extending direction of the slide rail 546 is parallel to the disposing direction of the syringe 534; one end of the sliding block 547 is fixedly connected with the reagent push rod 538, and the other end is slidably arranged on the sliding rail 546; the driving motor 548 can also be fixedly arranged on the protective box 535, and a motor shaft of the driving motor 548 is fixedly connected with the sliding block 547.

Therefore, when the gel in the gel storage device needs to be added into the reagent cylinder 537, the driving motor 548 drives the sliding block 547 to move upwards along the sliding rail 546, and the sliding block 547 can drive the reagent push rod 538 to move upwards in the reagent cylinder 537 so as to suck the gel; when the gel in the reagent cylinder 537 needs to be pushed into the capillary 505, the driving motor 548 drives the sliding block 547 to move down along the sliding rail 546, and the sliding block 547 can drive the reagent push rod 538 to move down in the reagent cylinder 537, so as to push out the gel.

The fluorescence detection device 600 includes a laser 601, a first optical path adjuster 602, a fixing bracket 603, an objective lens 604, a second optical path adjuster 605, and a spectrometer 606. The laser 601 is used to provide a light source, and the first optical path adjuster 602 is disposed at a front end of the laser 601, and includes a first plane mirror for emitting laser light emitted from the laser 601 to the left vertical direction, and a first lens for focusing the laser light emitted to the left vertical direction. In addition, a baffle 607 is fixed to the left side of the first optical path adjuster 602, the baffle 607 extends upward, and a through hole for passing the laser is provided on the baffle 607.

The fixing bracket 603 is disposed at the left side of the first optical path adjuster 602, the fixing bracket 603 is used to fix the capillary 505, the capillary 505 is in a laser focusing position, when the laser is applied to the capillary 505, the laser interacts with the fluorescent reagent in the capillary 505 to generate fluorescence, preferably, a connecting block 608 is fixed at the front side of the lower end of the baffle 607, and the fixing bracket 603 is fixed on the connecting block 608.

The objective lens 604 is located on the left side of the laser 601 and is parallel to the laser 601 and supported by an N-type holder 609, and the objective lens 604 is used for collecting the fluorescence emitted from the capillary 505. The second optical path adjuster 605 is disposed at the rear end of the objective lens 604, and includes a second planar mirror for changing the direction of the fluorescent light to the vertical upward, a filter for changing the fluorescent light to a monochromatic color in the vertical upward direction, and a second lens for focusing the monochromatic fluorescent light. The spectrometer 606 is disposed at the upper end of the second optical path adjuster 605, and the spectrometer 606 is used for performing spectrum analysis on the focused monochromatic fluorescence.

The integrated DNA analysis system integrates all steps of DNA detection and analysis on one device, is convenient to operate and quick to detect, and greatly improves the efficiency of DNA detection on the premise of ensuring the accuracy; the unique chip split design reduces the whole manufacturing difficulty and cost and improves the reaction effect on the premise of realizing the integrated function; the original micro-fluidic device realizes the fine control of the flow of the reaction fluid on the chip and ensures the smooth reaction on the chip by matching the fluid pipeline and the control valve which are designed in a layered way; the chip reaction system and the reagent supply system which are respectively and independently designed can meet the detection requirements of different quantities, and the flexibility of equipment use is improved; the integrated capillary electrophoresis system reduces the difficulty of replacing the capillary, improves the reaction efficiency and is convenient for non-professional personnel to use in daily life.

The present invention has been further described with reference to specific embodiments, but it should be understood that the detailed description should not be construed as limiting the spirit and scope of the present invention, and various modifications made to the above-described embodiments by those of ordinary skill in the art after reading this specification are within the scope of the present invention.

Claims

1. An integrated DNA analysis system, comprising:

a reagent supply system comprising a reagent storage device and a reagent injection device, the reagent storage device being in fluid communication with the reagent injection device;

the microfluidic reaction system comprises a microfluidic chip and a microfluidic device, wherein a reagent injection device is in fluid communication with the microfluidic chip, the microfluidic chip is connected with the microfluidic device, a sample to be analyzed can be added onto the microfluidic chip, a reaction reagent in the reagent storage device can be injected into the microfluidic chip by the reagent injection device, and the microfluidic device can control the flow of fluid in the microfluidic chip;

the micro-fluidic chip comprises a chip body and a reaction unit, wherein the chip body and the reaction unit are two independent split parts, and the formed chip body and the reaction unit are combined together to form the micro-fluidic chip; the chip body is provided with a sample extraction unit and a fluid pipeline, and the sample extraction unit is used for extracting reactants; the reaction unit comprises a reaction sheet, the reaction sheet is attached to the chip body, a reaction area is formed on the reaction sheet, the reaction area on the reaction sheet is communicated with a fluid pipeline on the chip body, and reactants and reaction reagents can be conveyed into the reaction area through the fluid pipeline so as to complete reaction in the reaction area;

the chip body comprises an accessory piece, a pipeline piece, a channel piece and a valve hole piece which are arranged in an overlapped mode, a sample extraction accessory is arranged on the accessory piece, a fluid pipeline is arranged on the pipeline piece, the channel piece is located below the pipeline piece and provides transitional connection for the communication of the fluid pipeline on the pipeline piece unit, and the valve hole piece is located below the channel piece and is used for the communication of the fluid pipeline on the pipeline piece; a pipeline disconnection point is arranged on a fluid pipeline of the pipeline piece, a transitional connection hole group is formed on the channel piece and corresponds to the pipeline disconnection point, a valve hole is formed on the valve hole piece and is communicated with the transitional connection hole group, and the fluid pipeline on the pipeline piece can be communicated with the pipeline disconnection point through the transitional connection hole group and the valve hole;

a reaction area connecting hole is formed on the channel sheet, a reactant inlet and a reactant outlet are formed on the reaction sheet, the reaction area on the reaction sheet is communicated with the corresponding reaction area connecting hole on the chip body through the reactant inlet, and reactants flow into the reaction area through the fluid pipeline, the reaction area connecting hole and the reactant inlet; the product after reaction can be conveyed to the corresponding reaction area connecting hole on the chip body through a reactant outlet on the reaction sheet;

the separation detection system comprises a capillary electrophoresis separation device and a fluorescence detection device, wherein the microfluidic chip is in fluid communication with the capillary electrophoresis separation device, the fluorescence detection device is connected with the capillary electrophoresis separation device, reactants on the microfluidic chip can enter the capillary electrophoresis separation device, and the fluorescence detection device can detect a separation result in the capillary electrophoresis separation device.

2. The integrated DNA analysis system according to claim 1, wherein the reagent storage means comprises a conventional reagent kit and a temperature control reagent kit, the conventional reagent kit comprises a reagent bottle capable of storing a reagent having a low requirement on the ambient temperature; the temperature control kit comprises a reagent bottle and a temperature control device, wherein the temperature control device can adjust the temperature of the reagent in the reagent bottle and can store the reagent with high requirement on the environmental temperature.

3. The integrated DNA analysis system of claim 2, wherein the temperature control kit comprises a heat-insulating sheath, the reagent bottle is arranged in the heat-insulating sheath, the temperature control component is arranged on the heat-insulating sheath, and the reagent bottle, the heat-insulating sheath and the temperature control component are arranged in the protective box body.

4. The integrated DNA analysis system according to claim 1, wherein the reagent injection device comprises a reagent accommodating chamber and an automatic pushing device, a reagent inlet of the reagent accommodating chamber is in fluid communication with a reagent outlet of the reagent storage device, and a reagent outlet of the reagent accommodating chamber is in fluid communication with a reagent inlet of the microfluidic chip; the automatic pushing device is connected with the reagent containing chamber, and can add the reagent in the reagent storage device to the reagent containing chamber and fill the reagent in the reagent containing chamber to the microfluidic chip.

5. The integrated DNA analysis system of claim 4, wherein the reagent injection device comprises a plurality of syringes vertically arranged side by side, each syringe is connected with a reagent push rod, each reagent push rod is respectively connected with a power assembly, and the power assemblies can respectively drive the pushing assemblies to reciprocate in the syringes so as to inject the reagents into the microfluidic chip in a classified manner.

6. The integrated DNA analysis system of claim 1, wherein the control valve plate unit is provided with an opening and closing valve at a position corresponding to the pipeline connecting hole, and the opening and closing valve can control the opening and closing of the pipeline connecting hole so as to realize the connection and disconnection of the corresponding fluid pipeline on the pipeline plate unit.

7. The integrated DNA analysis system of claim 1, wherein the microfluidic device comprises a chip carrier assembly and a connection hole opening and closing assembly, the microfluidic chip can be placed on the chip carrier assembly, the connection hole opening and closing assembly is connected to the pipeline connection hole in the chip placed on the chip carrier assembly, and the opening and closing of the pipeline connection hole in the chip can be controlled to connect and disconnect the fluid pipeline connected to the pipeline connection hole on the chip.

8. The integrated DNA analysis system of claim 7, wherein the temperature control assembly is disposed on the chip carrier assembly, and the temperature control assembly is in contact with the microfluidic chip after the microfluidic chip is placed on the chip carrier assembly, so as to adjust the reaction temperature on the microfluidic chip.

9. The integrated DNA analysis system according to claim 7, wherein the microfluidic device comprises a mechanical control assembly, and the mechanical control assembly is connected with the chip carrier assembly and can drive the chip carrier assembly to reciprocate.

10. The integrated DNA analysis system according to claim 1, wherein the capillary electrophoresis separating device comprises a capillary electrophoresis module and a temperature control module, the capillary electrophoresis module comprises a protection module, and the capillary is disposed inside the protection module; the temperature control assembly comprises an isolation assembly, and the heating assembly is arranged inside the isolation assembly; the capillary electrophoresis component and the temperature control component are in mutual contact and can perform heat transfer, and the heating component in the temperature control component can perform temperature regulation on the capillary in the capillary electrophoresis component.

11. The integrated DNA analysis system according to claim 10, wherein a detection window is provided on the guard member at a position corresponding to the fluid outlet end of the capillary, through which the electrophoretic separation in the capillary can be detected.

12. The integrated DNA analysis system of claim 10, wherein the capillary electrophoresis separation device comprises a quick-lock assembly, the quick-lock assembly being connected to the capillary electrophoresis assembly to removably dispose the capillary electrophoresis assembly in the integrated DNA analysis system.

13. The integrated DNA analysis system according to claim 10, wherein the capillary electrophoresis separating means comprises a gel advancing means comprising a gel receiving chamber for temporarily storing the gel, a temperature adjusting assembly connected to the gel receiving chamber for adjusting the temperature of the gel in the gel receiving chamber, and an automatic pushing means connected to the gel receiving chamber for adding the gel into the gel receiving chamber and pushing the gel in the gel receiving chamber to the capillary electrophoresis assembly.

14. The integrated DNA analysis system according to claim 13, wherein the gel propelling device comprises a reagent cylinder and a reagent pushing rod, a protection box is arranged outside the reagent cylinder, a semiconductor refrigerator is arranged in the protection box, the semiconductor refrigerator is connected with the reagent cylinder and can adjust the temperature of the gel in the reagent cylinder, the reagent pushing rod is connected with a propelling assembly, and the propelling assembly can drive the reagent pushing rod to reciprocate in the reagent cylinder.

15. The integrated DNA analysis system according to claim 1, wherein the fluorescence detection device comprises a laser, an objective lens and a spectrometer, the laser, the objective lens and the spectrometer are connected in sequence by optical paths, a first optical path adjuster is arranged between the laser and a capillary tube in the capillary electrophoresis separation device, the first optical path adjuster can change the path of the laser emitted by the laser, a second optical path adjuster is arranged between the objective lens and the spectrometer, and the second optical path adjuster can change the path of the fluorescence between the objective lens and the spectrometer.

16. The integrated DNA analysis system according to claim 15, wherein the first optical path adjuster includes a first plane mirror for reflecting the laser light emitted from the laser device perpendicularly toward the capillary direction and a first lens for focusing the laser light reflected toward the capillary direction.

17. The integrated DNA analysis system according to claim 15, wherein the second optical path adjuster comprises a second plane mirror, a filter and a second lens, the second plane mirror vertically reflecting the fluorescence emitted from the capillary tube toward the spectrometer; the filter can change the fluorescence reflected towards the spectrometer into a single color; the second lens may focus the monochromatic fluorescent light.