CN114774259B - Multichannel integrated rapid nucleic acid detection system and detection method - Google Patents
Multichannel integrated rapid nucleic acid detection system and detection method Download PDFInfo
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- CN114774259B CN114774259B CN202210479090.1A CN202210479090A CN114774259B CN 114774259 B CN114774259 B CN 114774259B CN 202210479090 A CN202210479090 A CN 202210479090A CN 114774259 B CN114774259 B CN 114774259B
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- 230000003321 amplification Effects 0.000 claims abstract description 14
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- 239000004065 semiconductor Substances 0.000 claims description 20
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/02—Burettes; Pipettes
- B01L3/021—Pipettes, i.e. with only one conduit for withdrawing and redistributing liquids
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L9/00—Supporting devices; Holding devices
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention provides a multichannel integrated nucleic acid rapid detection system which mainly comprises a motion control assembly, a temperature control assembly, a fluorescence scanning detection assembly, a working platform assembly and other structural assemblies. The system controls the integrated nucleic acid detection card box through the magnetic rod, the magnetic rod sleeve and the pressing rod, and is matched with the temperature control assembly and the fluorescence scanning detection assembly to automatically complete the nucleic acid extraction by the magnetic bead method, the configuration of an amplification detection system and the fluorescence amplification detection operation, and a detection report is automatically generated. The system can load 1-4 cartridges for single detection, and the single cartridge can realize multiple detections. The invention can realize multichannel integrated nucleic acid rapid detection, has simple operation, does not need manual intervention, can effectively improve the nucleic acid detection speed, and realizes the on-site rapid detection of pathogens.
Description
Technical Field
The invention belongs to the field of molecular diagnosis and medical detection, and particularly relates to a multichannel integrated rapid nucleic acid detection system and a detection method.
Background
The global pandemic of new crown epidemics has been over two years, but has not yet been effectively controlled. At present, the nucleic acid detection technology is still the gold standard for detecting the new coronavirus. Generally, the whole process of nucleic acid detection (including steps of nucleic acid extraction and purification, PCR amplification, detection and the like) needs to be completed by matching with various different instruments, and manual intervention is needed to transfer samples among different instruments in the detection process, so that the nucleic acid detection time is increased, the probability of cross contamination of samples is greatly increased, and the accuracy of detection results is influenced. Meanwhile, a large number of domestic basic medical institutions do not have perfect environmental conditions and equipment conditions to ensure that the basic medical institutions can independently complete nucleic acid detection. Therefore, the integrated nucleic acid detection technology has been regarded and developed in recent years, and the integrated nucleic acid detection system integrates all steps of nucleic acid detection into one system, so that the detection process does not need manual operation, and realizes sample input and result output. The integrated nucleic acid detection system does not need a high-specification laboratory environment, can be conveniently deployed and detected in primary medical institutions or quarantine sites, and can quickly and effectively cope with outbreak epidemic situations. Some integrated nucleic acid detecting devices based on cartridges or microfluidic chips are available in the market, and can fully automatically realize the whole process of nucleic acid detection, such as GeneXpert series products of Sapei company and FilmArray series products of Merrieia company. However, most of the card boxes matched with the devices have complex structures and very high prices, and are not suitable for popularization and application in areas with poor medical resources, such as primary medical institutions and the like.
Disclosure of Invention
The purpose of the invention is as follows: the invention aims to provide a multichannel integrated rapid nucleic acid detection system, which is based on a simple, portable and efficient integrated card box, can realize multichannel integrated rapid nucleic acid detection, can realize sample input and result output, and does not need manual intervention in the whole detection process.
The technical scheme is as follows: the multichannel integrated rapid nucleic acid detection system comprises a motion control assembly, a temperature control assembly, a fluorescence scanning detection assembly, a working platform assembly, a base, a back plate and a switching power supply;
the motion control assembly comprises a power mechanism, a transmission mechanism and an execution mechanism; the power mechanism comprises a horizontal movement motor, a lower press rod control motor, a magnetic bar control motor and a magnetic bar sleeve control motor; the transmission mechanism comprises a horizontal movement module, a lower pressing rod movement module, a vertical sliding rail, a magnetic bar frame sliding block, a magnetic sleeve frame sliding block, a magnetic bar frame synchronous belt and a magnetic bar sleeve frame synchronous belt; the actuating mechanism comprises a magnetic bar rack and a magnetic bar, a magnetic bar sleeve rack and a magnetic bar sleeve, and a lower pressure bar rack and a lower pressure bar;
the horizontal movement module is fixedly arranged on the base, the horizontal movement motor is arranged on the right side of the horizontal movement module, the motor shaft is connected with the screw rod through the coupler, and the movement platform bottom plate is fixedly arranged on the screw rod sliding table; the lower pressure rod movement module is fixedly arranged on the right side of the vertical plate of the movement platform, the lower pressure rod control motor is arranged below the lower pressure rod movement module, and the lower pressure rod frame is fixedly arranged on a lead screw sliding table of the lower pressure rod movement module; the vertical slide rail is fixedly arranged on the left side of a vertical plate of the motion platform, the magnetic rod frame slide block and the magnetic rod sleeve frame slide block are arranged on the vertical slide rail in a sliding manner, the magnetic rod frame slide block is arranged above the magnetic rod frame slide block, and the magnetic rod sleeve frame slide block is arranged below the magnetic rod sleeve frame slide block; the magnetic rod frame is fixedly arranged on the magnetic rod frame sliding block, and the magnetic rod sleeve frame is fixedly arranged on the magnetic rod sleeve frame sliding block; one end of the magnetic bar frame is fixed with the magnetic bar frame synchronous belt through the pressing sheet, and one end of the magnetic bar sleeve frame is fixed with the magnetic bar sleeve frame synchronous belt through the pressing sheet;
the temperature control assembly adopts a multilayer structure design and is respectively as follows from bottom to top: the heat radiation structure is composed of 2 heat radiation fans and heat radiation fins; a heat conducting bottom plate and a heat insulating base; the heating structure consists of 4 semiconductor refrigerating plates and 1 aluminum-based resistance heating plate; and a heat conduction structure consisting of a No. 1 metal heat trap, a No. 2 metal heat trap and a No. 3 metal heat trap;
the fluorescence scanning detection assembly comprises a fluorescence scanning motor, a motor frame, a bearing frame, a synchronous wheel, a synchronous belt, a sliding rail supporting frame, a sliding rail, a sliding block, a fluorescence module base and a fluorescence module.
Further, the magnetic rod sleeve is inserted into and fixed on the magnetic rod sleeve frame; the magnetic rods are vertically arranged below the magnetic rod rack, each magnetic rod is vertically aligned with the corresponding magnetic rod sleeve, and the magnetic rods can extend into the corresponding magnetic rod sleeves; the lower pressure rod is vertically arranged below the lower pressure rod frame.
Further, fluorescence module fixed mounting passes through the screw fixation on the slider on the fluorescence module base, and the base protruding portion passes through the briquetting and fixes with the hold-in range, and the slider can follow slide rail horizontal movement.
Furthermore, the working platform assembly comprises a front support frame, a rear support frame, a left support frame, a working panel, a shading panel, a card box fixing pressing block, a card box fixing sliding block and 2 fixing knobs; 4 strip-shaped openings are formed in the middle of the working panel, and the card box is inserted into the lower temperature control assembly through the openings; one end of the opening is provided with a card box fixing pressing block, and the other end of the opening is provided with a card box fixing sliding block; after the card box is packed into, the left breach of the fixed briquetting of card box can support the card box left end, and the fixed slider of card box slides on the left and withholds the card box right-hand member to screw up fixed knob, can realize the fixed of card box.
Furthermore, the horizontal motion motor drives a screw rod in the horizontal motion module to rotate, so as to drive the motion platform to move in the horizontal direction, and therefore the horizontal motion motor controls all parts on the working platform to move left and right in the horizontal direction;
the lower pressure rod control motor drives a screw rod in the lower pressure rod movement module to rotate, so that the lower pressure rod frame is driven to move in the vertical direction, and the lower pressure rod is controlled by the lower pressure rod control motor to move up and down in the vertical direction;
the magnetic bar control motor rotates to drive the magnetic bar rack synchronous belt to move in a transmission mode in the vertical direction, so that the magnetic bar rack mounted on the magnetic bar rack sliding block is driven to move up and down along the vertical sliding rail, and the magnetic bar control motor controls the magnetic bar to move up and down in the vertical direction;
the bar magnet sleeve control motor rotates to drive the bar magnet sleeve frame synchronous belt to move in a transmission mode in the vertical direction, so that the bar magnet sleeve frame mounted on the bar magnet sleeve frame sliding block is driven to move up and down along the vertical sliding rail, and the bar magnet sleeve control motor controls the bar magnet sleeve to move up and down in the vertical direction.
Furthermore, temperature sensors are arranged in the No. 1 metal hot trap, the No. 2 metal hot trap and the No. 3 metal hot trap, and the temperature of the hot traps is monitored in real time and is used as a feedback signal for temperature control;
the aluminum-based resistance heating plate is positioned below the No. 1 metal hot trap and is used for heating the No. 1 metal hot trap; the 4 semiconductor refrigerating plates are divided into two groups and respectively positioned below the No. 2 metal hot trap and the No. 3 metal hot trap, and the No. 2 metal hot trap and the No. 3 metal hot trap can be heated and refrigerated;
the heat insulation base is positioned below the aluminum-based resistance heating plate, so that heat generated by the aluminum-based resistance heating plate is prevented from being conducted to the radiating fins below the aluminum-based resistance heating plate; the heat conducting plate is positioned below the semiconductor refrigerating sheet and conducts heat on the lower surface of the semiconductor to the radiating fins below the semiconductor refrigerating sheet, so that the temperature difference between the front side and the back side of the semiconductor refrigerating sheet is ensured to be within a working range; the heat radiation fan blows air to the heat radiation fins to promote heat radiation of the heat radiation fins.
Furthermore, the fluorescence scanning motor drives the synchronous belt to horizontally drive through the synchronous wheel, so that the fluorescence module is driven to move in the horizontal direction along the sliding rail, the fluorescence module lens sweeps through the amplification detection tube of the card box, the fluorescence module emits exciting light, excited fluorescence signals are collected and sent to the upper computer software for processing, and the fluorescence scanning detection function is achieved.
After the card box matched with the system design is loaded into the system, the operations of magnetic bead method nucleic acid extraction, amplification detection system configuration, multiple amplification detection and the like can be fully automatically completed by matching the system magnetic rod, the magnetic rod sleeve and the lower pressing rod with the temperature control assembly and the fluorescence scanning detection assembly, and finally, the upper computer software matched with the system can form a detection report after analysis and calculation according to the acquired fluorescence data.
Further, the multi-channel is not limited to the single test shown in the figure to load 4 cartridges at most, and 4 magnetic rods, 4 magnetic rod sleeves and 4 press rods are provided to realize the parallel driving control of the cartridges. Through horizontal extension, set up the card box installation position of more quantity promptly, increase bar magnet, bar magnet cover passageway and depression bar quantity down simultaneously, can realize loading and control the card box of more quantity in the single detects on this system, further improve and detect the flux. The invention also provides a detection method of the multichannel integrated nucleic acid rapid detection system, and the detection process comprises the following specific execution actions:
loading a card box: the reagent required for detection is pre-loaded in the corresponding hole site of the card box, the sample to be detected is added in the hole site No. 1, the card box is inserted through the opening on the working panel and is fixed by the card box fixing pressing block and the card box fixing sliding block, the detection can be started, and 1-4 card boxes can be loaded in one detection;
magnetic bead mixing, suction, transfer and release: the horizontal movement motor can control the magnetic rod, the magnetic rod sleeve and the lower pressing rod to move to the position above the hole position to be operated of the card box; the magnetic rod sleeve controls the motor to control the magnetic rod sleeve to downwards enter the liquid in the corresponding hole position of the card box, and controls the magnetic rod sleeve to vibrate up and down at a high speed, so that the action of uniformly mixing magnetic beads is realized; the magnetic rod control motor controls the magnetic rod to downwards enter the magnetic rod sleeve, then the magnetic rod control motor and the magnetic rod sleeve control motor simultaneously control the magnetic rod and the magnetic rod sleeve to downwards enter the liquid in the corresponding hole position of the card box, magnetic beads in the liquid can be adsorbed to the bottom of the magnetic sleeve, and magnetic bead absorption action is realized; the magnetic bar control motor and the magnetic bar sleeve control motor simultaneously control the magnetic bar and the magnetic bar sleeve to move upwards and leave the card box, then the horizontal movement motor controls the movement platform to move horizontally, so that the magnetic bar and the magnetic bar sleeve move to the position above a magnetic hole position of the card box, the magnetic bar control motor and the magnetic bar sleeve control motor simultaneously control the magnetic bar and the magnetic bar sleeve to move downwards into liquid at the magnetic hole position of the card box, the magnetic bar control motor controls the magnetic bar to rapidly ascend, the magnetic bar sleeve controls the motor to control the magnetic bar sleeve to vibrate at high speed, magnetic beads adsorbed at the bottom of the magnetic bar sleeve are released into the liquid, and the transfer and release actions of the magnetic beads are realized;
temperature control: the No. 1 metal heat trap takes an aluminum-based resistance heating plate as a heat source and can heat a cracking hole of the cartridge; the No. 2 metal hot trap takes 2 semiconductor refrigerating sheets as a heat source, and can heat and refrigerate the elution hole site of the card box; the No. 3 metal hot trap takes 2 semiconductor refrigerating sheets as heat sources, and can carry out thermal cycle control or constant temperature control on the card box amplification detection tube;
liquid transfer: the lower press rod control motor controls the lower press rod to move downwards, the lower press valve in the card box is pressed for a certain distance, so that channels of hole sites on two sides of the lower press valve can be opened, the lower press rod presses the lower press valve in the card box again, and the channels on two sides of the lower press valve can be closed; the lower pressing rod is controlled by the lower pressing rod control motor to move downwards to the position above the piston of the card box, the lower pressing rod is controlled by the horizontal movement motor to move leftwards for a small section, the combination of the lower pressing rod and the piston can be realized, then the lower pressing rod can drive the piston to move up and down, the piston can move upwards to suck liquid into the piston tube, the piston can push the liquid out of the piston tube downwards, the liquid can be repeatedly sucked and pushed out by the repeated up and down movement of the piston, and the uniform mixing of the liquid is realized; at this moment, the purified sample can be configured by controlling a lower pressure valve and a piston through a lower pressure rod, and is transferred to an amplification detection tube for amplification detection;
fluorescence scanning detection: at the specific stage of extension detection, fluorescence scanning motor control fluorescence module moves along the slide rail level, and fluorescence module camera lens passes through the augmentation detecting tube of every card box in proper order, and fluorescence module sends the exciting light simultaneously to gather the fluorescence signal of arousing, and convert the light signal into the signal of telecommunication, send for host computer software and handle.
Has the advantages that: compared with the prior art, the invention has the following remarkable advantages: the multichannel integrated rapid nucleic acid detection system can load a plurality of card boxes in single detection, and controls the plurality of card boxes in parallel to complete nucleic acid extraction, amplification detection system configuration and multiplex amplification detection by matching the magnetic rod, the magnetic rod sleeve and the lower pressing rod with the temperature control assembly and the fluorescence scanning detection assembly, thereby realizing sample input and result output. The detection process does not need manual intervention, and the method is simple to operate, convenient and quick. In addition, through horizontal extension, set up more quantity card box installation positions, increase bar magnet, bar magnet cover passageway and depression bar quantity down simultaneously, can further improve the detection flux very conveniently.
Drawings
FIG. 1 is a front elevational view of the system as a whole;
FIG. 2 is a block diagram of a motion control assembly;
FIG. 3 is a front view block diagram of a temperature control assembly;
FIG. 4 is a diagram of a fluorescence scanning detection assembly;
FIG. 5 is a diagram of a work platform assembly;
figure 6 is a front and back schematic view of the combination of the depressible bar and the cartridge piston.
Detailed Description
The technical scheme of the invention is further explained by combining the attached drawings.
As shown in fig. 1, the system as a whole includes: the device comprises a motion control component 1, a temperature control component 2, a fluorescence scanning detection component 3, a working platform component 4, a base 5, a back plate 6, a switching power supply 7, a card box 8 loaded in the switching power supply and the like; the temperature control component 2 and the fluorescence scanning detection component 3 are positioned below the working platform component 4, the fluorescence scanning detection component 3 is positioned on the right side of the temperature control component 2, and the card box 8 is arranged in the temperature control component 2 through a mounting hole of the working platform component 4; the motion control assembly 1 is positioned behind the working platform assembly 4, and a magnetic bar frame 112, a magnetic bar sleeve frame 110 and a lower pressure bar frame 116 in the motion control assembly 1 extend forwards and are positioned above the working platform assembly 4 to control the installed card box 8; the back plate 6 and the switching power supply 7 are positioned behind the motion control component 1;
as shown in fig. 2, the motion control assembly 1 includes a horizontal motion motor 101, a lower press rod control motor 102, a magnetic rod control motor 103, a magnetic rod sleeve control motor 104, a magnetic sleeve frame slide block 105, a vertical slide rail 106, a horizontal motion module 107, a motion platform base plate 108, a magnetic rod sleeve frame synchronous belt 109, a magnetic rod sleeve frame 110, a magnetic rod frame synchronous belt 111, a magnetic rod frame 112, a motion platform vertical plate 113, a magnetic rod frame slide block 114, a lower press rod motion module 115, a lower press rod frame 116, a magnetic rod 117, a lower press rod 118, and a magnetic rod sleeve 119; the horizontal movement module 107 is fixedly arranged on the base 5, the horizontal movement motor 101 is arranged on the right side of the horizontal movement module 107, a motor shaft is connected with a screw rod through a coupler, and the movement platform bottom plate 108 is fixedly arranged on a screw rod sliding table; the lower pressure rod movement module 115 is fixedly arranged on the right side of the vertical plate 113 of the movement platform, the lower pressure rod control motor 102 is arranged below the lower pressure rod movement module 115, and the lower pressure rod frame 116 is fixedly arranged on a screw rod sliding table of the lower pressure rod movement module 115; the vertical slide rail 106 is fixedly arranged on the left side of a vertical plate 113 of the motion platform, the magnetic bar frame slide block 114 and the magnetic bar sleeve frame slide block 105 are arranged on the vertical slide rail 106 in a sliding manner, the magnetic bar frame slide block 114 is arranged above the magnetic bar frame slide block, and the magnetic bar sleeve frame slide block 105 is arranged below the magnetic bar frame slide block; the magnetic rod frame 112 is fixedly arranged on the magnetic rod frame sliding block 114, and the magnetic rod sleeve frame 110 is fixedly arranged on the magnetic rod sleeve frame sliding block 105; one end of a magnetic rod frame 112 is fixed with a magnetic rod frame synchronous belt 111 through a pressing sheet, and one end of a magnetic rod sleeve frame 110 is fixed with a magnetic rod sleeve frame synchronous belt 109 through a pressing sheet; the magnetic rod sleeve 119 can be inserted into and fixed on the magnetic rod sleeve frame 110; the magnetic rods 117 are vertically arranged below the magnetic rod rack 112, each magnetic rod 117 is vertically aligned with the corresponding magnetic rod sleeve 119, and the magnetic rods 117 can extend into the corresponding magnetic rod sleeves 119; the lower pressure lever 118 is vertically installed below the lower pressure lever bracket 116; under the cooperation of the horizontal movement motor 101, the magnetic rod control motor 103, the magnetic rod sleeve control motor 104 and the lower press rod control motor 102, the magnetic rod 117, the magnetic rod sleeve 119 and the lower press rod 118 can be controlled to move in the horizontal and vertical directions respectively.
As shown in fig. 3, the temperature control assembly 2 is designed in a multi-layer structure, which is from bottom to top: a heat dissipation structure composed of 2 heat dissipation fans 201 and heat dissipation fins 202; a thermally conductive base plate 203 and a thermally insulating base 208; the heating structure consists of 4 semiconductor refrigerating fins 204 and 1 aluminum-based resistance heating plate; and a heat conducting structure consisting of a metal heat trap No. 1 207, a metal heat trap No. 2 206, and a metal heat trap No. 3 205.
As shown in fig. 4, the fluorescence detection assembly 3 includes a fluorescence scanning motor 301, a motor frame 302, a timing belt 303, a bearing frame 304, a synchronizing wheel 305, a rail support frame 306, a rail 307, a sliding block 308, a fluorescence module base 309, a fluorescence module 310, wherein the fluorescence module 310 is fixedly mounted on the fluorescence module base 309, the base is fixed on the sliding block 308 through a screw, a protruding portion of the base is fixed with the timing belt 303 through a pressing block, and the sliding block 308 can horizontally move along the sliding rail 307 under the driving of the fluorescence scanning motor 301.
As shown in fig. 5, the work platform assembly comprises a front support frame 401, a rear support frame 402, a shading panel 403, a cartridge fixing slider 404, a fixing knob 405, a work panel 406, a cartridge fixing pressing block 407, a left support frame 408, 4 elongated openings in the middle of the work panel 406, and a cartridge 8 inserted into the lower temperature control assembly 2 through the openings; a card box fixing pressing block 407 is arranged at one end of the opening, and a card box fixing sliding block 404 is arranged at the other end of the opening; after the card box 8 is installed, the left notch of the card box fixing pressing block 407 can abut against the left end of the card box, the left sliding of the card box fixing sliding block 404 presses the right end of the card box, and the fixing knob 405 is screwed down, so that the fixation of the card box can be realized.
As shown in fig. 6, which is a schematic position diagram of the push rod 118 and the cartridge piston 801 before (a) and after (b), after the combination, the push rod 118 can push and pull the cartridge piston 801 to suck and release liquid.
Examples
The whole nucleic acid detection process comprises magnetic bead transfer-based nucleic acid extraction by a magnetic bead method, PCR amplification system configuration and real-time fluorescence PCR amplification detection, and an integrated nucleic acid detection card box designed in cooperation with the system is required to be used in the embodiment.
Firstly, add the sample that awaits measuring in the schizolysis hole site in the card box of having preloaded detect reagent to insert the card box to the specific station of temperature control assembly through the opening of work panel, the fixed briquetting of card box blocks and fixes the card box left end, moves the fixed slider of card box left and pushes down the card box right-hand member, and screws up fixed knob, fixes the card box.
The horizontal motion motor controls the magnetic rod and the magnetic rod sleeve to move above the magnetic bead hole of the card box, the magnetic rod control motor and the magnetic rod sleeve control motor control the magnetic rod and the magnetic rod sleeve to simultaneously descend into the magnetic bead liquid, and the magnetic bead is adsorbed to the bottom of the magnetic rod sleeve; a magnetic rod control motor and a magnetic rod sleeve control motor control a magnetic rod and a magnetic rod sleeve to lift away from a magnetic bead hole of a card box, a horizontal movement motor controls the magnetic rod and the magnetic rod sleeve to move above the cracking hole of the card box, the magnetic rod control motor and the magnetic rod sleeve control motor control the magnetic rod and the magnetic rod sleeve to simultaneously descend into a cracking solution, the magnetic rod control motor controls the magnetic rod to rapidly lift away from the cracking hole, the magnetic rod sleeve control motor controls the magnetic rod sleeve to vibrate up and down at a high speed, and magnetic beads are released and uniformly mixed; meanwhile, the resistance heating plate heats the liquid in the cracking hole through a No. 1 metal hot trap according to the set cracking temperature, so as to promote the cracking of the sample in the cracking hole;
after the cracking is finished, the magnetic rod and the magnetic rod sleeve repeat the actions of adsorbing magnetic beads and transferring magnetic beads, the magnetic beads are transferred to a washing hole position from a cracking hole position, the magnetic rod sleeve controls a motor to control the magnetic rod sleeve to vibrate up and down at a high speed, the magnetic beads are released and uniformly mixed, and the first washing is finished;
repeating the actions, transferring the magnetic beads from the No. 1 washing hole site to the No. 2 washing hole site, and uniformly mixing the magnetic beads by high-speed vibration to complete secondary washing;
after washing is finished, repeating the actions of adsorbing magnetic beads and transferring magnetic beads, transferring the magnetic beads from the No. 2 washing hole position to an alcohol removing hole position, ensuring that the magnetic rod does not rise, and controlling the motor by the magnetic rod control motor and the magnetic rod sleeve to control the magnetic rod and the magnetic rod sleeve to move up and down slowly for multiple times at the same time so as to remove alcohol impurities remained on the surfaces of the magnetic beads;
after the alcohol is removed, the magnetic bar and the magnetic bar sleeve act together to transfer the magnetic beads to an elution hole, the magnetic bar quickly rises away from the elution hole, the magnetic bar vibrates at a high speed and uniformly mixes, and the magnetic beads are released and uniformly mixed; meanwhile, the semiconductor refrigerating sheet heats the liquid in the elution hole through a No. 2 metal hot trap according to the set elution temperature, so that the purified sample is promoted to be separated from the magnetic beads and enter the eluent;
after elution is finished, refrigerating the semiconductor refrigerating sheet, and cooling the eluent by a No. 2 metal hot trap;
the horizontal motion motor controls the lower pressure rod to move to the position above a No. 1 lower pressure valve, the lower pressure rod controls the motor to control the lower pressure rod to press down the No. 1 lower pressure valve so as to open a channel between an elution hole and a piston pipe, the horizontal motion motor controls the lower pressure rod to move to the position above the piston, the lower pressure rod controls the motor to control the lower pressure rod to approach the piston, the horizontal motion motor moves the lower pressure rod to the left for a short distance so as to complete the combination of the lower pressure rod and the piston, then the lower pressure rod controls the motor to control the lower pressure rod to move up and down repeatedly so that eluent enters the piston pipe, PCR solid freeze-drying reagents preloaded in the piston pipe are dissolved and mixed uniformly, finally the lower pressure rod is controlled to keep the piston in a suction state, liquid stays in the piston pipe, the horizontal motion motor moves the lower pressure rod to the right for a short distance so as to complete the separation of the lower pressure rod and the piston, then the lower pressure rod controls the motor to lift the lower pressure rod to leave the piston pipe, the piston is controlled by the horizontal motion motor to return to the position above the No. 1 lower pressure rod, the lower pressure rod controls the lower pressure rod to press down the valve again so as to close the channel between the elution hole and the piston pipe; the lower pressure rod is controlled by a horizontal movement motor to move right to the position above the No. 2 lower pressure valve, the lower pressure rod is controlled by a lower pressure rod control motor to press the No. 2 lower pressure valve to open a channel between the piston tube and the PCR diffusion detection tube, and then the lower pressure rod is controlled by a lower pressure rod control motor to ascend to the position above the card box; the horizontal movement motor controls the lower pressure rod to move back to the upper side of the piston, the lower pressure rod controls the motor to control the lower pressure rod to press the piston, liquid in the piston tube is pushed into the PCR amplification detection tube, and then the lower pressure rod controls the motor to control the lower pressure rod to ascend to the upper side of the card box; the horizontal movement motor controls the lower pressure rod to move right to the position above the No. 2 lower pressure valve, the lower pressure rod controls the motor to control the lower pressure rod to press down the No. 2 lower pressure valve again to close a channel between the piston tube and the PCR amplification detection tube, and therefore the PCR system configuration is completed;
then, the semiconductor refrigerating plate controls the liquid temperature in the PCR reaction tubes through a No. 3 metal hot trap according to the thermal cycle temperature set by a program, and controls a fluorescence module to transversely scan each PCR reaction tube through a fluorescence scanning motor at a specific detection stage, wherein the fluorescence module emits exciting light, collects excited fluorescence signals, converts the optical signals into electric signals and sends the electric signals to upper computer software for processing and displaying;
and finally, generating a detection report by analyzing the fluorescence curve, wherein the whole process does not need manual intervention, and realizes sample input and result output.
Claims (8)
1. A multi-channel integrated rapid nucleic acid detection system is characterized by comprising a motion control assembly, a temperature control assembly, a fluorescence scanning detection assembly, a working platform assembly, a base, a back plate and a switching power supply;
the motion control assembly comprises a power mechanism, a transmission mechanism and an execution mechanism; the power mechanism comprises a horizontal movement motor, a lower press rod control motor, a magnetic bar control motor and a magnetic bar sleeve control motor; the transmission mechanism comprises a horizontal movement module, a lower pressing rod movement module, a vertical sliding rail, a magnetic bar frame sliding block, a magnetic sleeve frame sliding block, a magnetic bar frame synchronous belt and a magnetic bar sleeve frame synchronous belt; the actuating mechanism comprises a magnetic bar rack and a magnetic bar, a magnetic bar sleeve rack and a magnetic bar sleeve, and a lower pressure bar rack and a lower pressure bar;
the horizontal movement module is fixedly arranged on the base, the horizontal movement motor is arranged on the right side of the horizontal movement module, the motor shaft is connected with the screw rod through the coupler, and the movement platform bottom plate is fixedly arranged on the screw rod sliding table; the lower pressure rod movement module is fixedly arranged on the right side of the vertical plate of the movement platform, the lower pressure rod control motor is arranged below the lower pressure rod movement module, and the lower pressure rod frame is fixedly arranged on a lead screw sliding table of the lower pressure rod movement module; the vertical sliding rail is fixedly arranged on the left side of the vertical plate of the motion platform, the magnetic bar frame sliding block and the magnetic bar sleeve frame sliding block are arranged on the vertical sliding rail in a sliding mode, the magnetic bar frame sliding block is arranged above the magnetic bar frame sliding block, and the magnetic bar sleeve frame sliding block is arranged below the magnetic bar frame sliding block; the magnetic bar frame is fixedly arranged on the magnetic bar frame sliding block, and the magnetic bar sleeve frame is fixedly arranged on the magnetic bar sleeve frame sliding block; one end of the magnetic bar frame is fixed with the magnetic bar frame synchronous belt through the pressing sheet, and one end of the magnetic bar sleeve frame is fixed with the magnetic bar sleeve frame synchronous belt through the pressing sheet;
the temperature control assembly is designed by adopting a multilayer structure and comprises the following components from bottom to top: the heat radiation structure is composed of 2 heat radiation fans and heat radiation fins; a heat conducting bottom plate and a heat insulating base; the heating structure consists of 4 semiconductor refrigerating plates and 1 aluminum-based resistance heating plate; and a heat conduction structure consisting of a No. 1 metal heat trap, a No. 2 metal heat trap and a No. 3 metal heat trap;
the fluorescence scanning detection assembly comprises a fluorescence scanning motor, a motor frame, a bearing frame, a synchronous wheel, a synchronous belt, a sliding rail supporting frame, a sliding rail, a sliding block, a fluorescence module base and a fluorescence module;
the working platform assembly comprises a front support frame, a rear support frame, a left support frame, a working panel, a shading panel, a card box fixing pressing block, a card box fixing sliding block and 2 fixing knobs; 4 strip-shaped openings are formed in the middle of the working panel, and the card box is inserted into the lower temperature control assembly through the openings; one end of the opening is provided with a card box fixing pressing block, and the other end of the opening is provided with a card box fixing sliding block; after the card box is arranged, a gap on the left side of the card box fixing pressing block can abut against the left end of the card box, the right end of the card box is pressed by the left sliding of the card box fixing sliding block, and the fixing knob is screwed down to fix the card box;
the horizontal motion motor drives a screw rod in the horizontal motion module to rotate so as to drive the motion platform to move in the horizontal direction, so that the horizontal motion motor controls all parts on the working platform to move left and right in the horizontal direction;
the lower pressure rod control motor drives a screw rod in the lower pressure rod movement module to rotate, so that the lower pressure rod frame is driven to move in the vertical direction, and the lower pressure rod is controlled by the lower pressure rod control motor to move up and down in the vertical direction;
the utility model discloses a liquid mixing box, including box piston, depression bar control motor control depression bar downstream to card box piston top, the depression bar moves a segment left under the horizontal motion motor control, can realize depression bar and piston combination down, and then the depression bar can drive piston up-and-down motion down, and piston up-and-down motion can be inhaled liquid to the piston pipe, and the piston can be pushed liquid piston pipe downwards, and the piston up-and-down motion can be inhaled and release liquid repeatedly, realizes the liquid mixing.
2. The multi-channel integrated rapid nucleic acid detection system according to claim 1, wherein the magnetic rod sleeve is inserted into and fixed on the magnetic rod sleeve frame; the magnetic rods are vertically arranged below the magnetic rod rack, each magnetic rod is vertically aligned with the corresponding magnetic rod sleeve, and the magnetic rods can extend into the corresponding magnetic rod sleeves; the lower pressure rod is vertically arranged below the lower pressure rod frame.
3. The multi-channel integrated rapid nucleic acid detection system of claim 1, wherein the fluorescence module is fixedly mounted on a fluorescence module base, the base is fixed on a slide block through a screw, a base protruding part is fixed with a synchronous belt through a pressing block, and the slide block can horizontally move along a slide rail.
4. The multi-channel integrated rapid nucleic acid detection system according to claim 1,
the magnetic bar control motor rotates to drive the magnetic bar rack synchronous belt to move in a transmission mode in the vertical direction, so that the magnetic bar rack arranged on the magnetic bar rack sliding block is driven to move up and down along the vertical sliding rail, and the magnetic bar control motor controls the magnetic bar to move up and down in the vertical direction;
the bar magnet sleeve control motor rotates to drive the bar magnet sleeve frame synchronous belt to move in a transmission mode in the vertical direction, so that the bar magnet sleeve frame mounted on the bar magnet sleeve frame sliding block is driven to move up and down along the vertical sliding rail, and the bar magnet sleeve control motor controls the bar magnet sleeve to move up and down in the vertical direction.
5. The multi-channel integrated rapid nucleic acid detection system according to claim 1, wherein temperature sensors are installed inside the metal heat trap No. 1, the metal heat trap No. 2 and the metal heat trap No. 3, and the temperature of the heat traps is monitored in real time and is used as a feedback signal for temperature control;
the aluminum-based resistance heating plate is positioned below the No. 1 metal hot trap and is used for heating the No. 1 metal hot trap; the 4 semiconductor refrigerating plates are divided into two groups and are respectively positioned below the No. 2 metal hot trap and the No. 3 metal hot trap, and the No. 2 metal hot trap and the No. 3 metal hot trap can be heated and refrigerated;
the heat insulation base is positioned below the aluminum-based resistance heating plate, so that heat generated by the aluminum-based resistance heating plate is prevented from being conducted to the radiating fins below the aluminum-based resistance heating plate; the heat conduction bottom plate is positioned below the semiconductor refrigerating sheet, and is used for conducting heat on the lower surface of the semiconductor to the radiating fins below the semiconductor refrigerating sheet, so that the temperature difference between the front surface and the back surface of the semiconductor refrigerating sheet is within a working range; the heat radiation fan blows air to the heat radiation fins to promote heat radiation of the heat radiation fins.
6. The multi-channel integrated rapid nucleic acid detection system of claim 1, wherein the fluorescence scanning motor drives the synchronous belt to horizontally drive through the synchronous wheel, so as to drive the fluorescence module to move along the slide rail in the horizontal direction, the fluorescence module lens sweeps across the amplification detection tube of the card box, the fluorescence module emits excitation light, collects the excited fluorescence signal, and sends the fluorescence signal to the upper computer for software processing, so as to realize the fluorescence scanning detection function.
7. The multi-channel integrated rapid nucleic acid detection system according to claim 1, wherein the multi-channel integrated rapid nucleic acid detection system is capable of being expanded laterally, and is provided with a greater number of cartridge installation sites, each multi-channel is loaded with a maximum of 4 cartridges, and is provided with 4 magnetic rods, 4 magnetic rod sleeves and 4 push rods for parallel driving control of the cartridges.
8. The detection method of the multi-channel integrated rapid nucleic acid detection system according to claim 1, wherein the detection process comprises the following specific actions:
loading a card box: the reagent required for detection is pre-loaded in the corresponding hole site of the card box, the sample to be detected is added in the hole site No. 1, the card box is inserted through the opening on the working panel and is fixed by the card box fixing pressing block and the card box fixing sliding block, the detection can be started, and 1-4 card boxes can be loaded in one detection;
magnetic bead mixing, suction, transfer and release: the horizontal movement motor can control the magnetic rod, the magnetic rod sleeve and the lower pressing rod to move to the position above the hole position to be operated of the card box; the magnetic rod sleeve controls the motor to control the magnetic rod sleeve to enter the liquid in the corresponding hole position of the card box downwards, and controls the magnetic rod sleeve to vibrate up and down at a high speed, so that the uniform mixing action of magnetic beads is realized; the magnetic rod control motor controls the magnetic rod to downwards enter the magnetic rod sleeve, then the magnetic rod control motor and the magnetic rod sleeve control motor simultaneously control the magnetic rod and the magnetic rod sleeve to downwards enter liquid of corresponding hole sites of the card box, magnetic beads in the liquid can be adsorbed to the bottom of the magnetic sleeve, and magnetic bead absorption action is realized; the magnetic bar control motor and the magnetic bar sleeve control motor simultaneously control the magnetic bar and the magnetic bar sleeve to move upwards and leave the card box, then the horizontal movement motor controls the movement platform to move horizontally, so that the magnetic bar and the magnetic bar sleeve move to the position above a magnetic hole position of the card box, the magnetic bar control motor and the magnetic bar sleeve control motor simultaneously control the magnetic bar and the magnetic bar sleeve to move downwards into liquid at the magnetic hole position of the card box, the magnetic bar control motor controls the magnetic bar to rapidly ascend, the magnetic bar sleeve controls the motor to control the magnetic bar sleeve to vibrate at high speed, magnetic beads adsorbed at the bottom of the magnetic bar sleeve are released into the liquid, and the transfer and release actions of the magnetic beads are realized;
temperature control: the No. 1 metal heat trap takes an aluminum-based resistance heating plate as a heat source and can heat a cracking hole of the cartridge; the No. 2 metal hot trap takes 2 semiconductor refrigerating sheets as a heat source, and can heat and refrigerate the elution hole site of the card box; the No. 3 metal hot trap takes 2 semiconductor refrigerating sheets as heat sources, and can carry out thermal cycle control or constant temperature control on the card box amplification detection tube;
liquid transfer: the lower pressing rod controls the motor to control the lower pressing rod to move downwards, a lower pressing valve in the card box is pressed for a certain distance, so that channels of hole sites on two sides of the lower pressing valve can be opened, the lower pressing rod presses the lower pressing valve in the card box again, and the channels on two sides of the lower pressing valve can be closed; the lower pressing rod is controlled by the lower pressing rod control motor to move downwards to the position above the piston of the card box, the lower pressing rod is controlled by the horizontal movement motor to move leftwards for a small section, the combination of the lower pressing rod and the piston can be realized, then the lower pressing rod can drive the piston to move up and down, the piston can suck liquid into the piston tube by moving upwards, the piston can push the liquid out of the piston tube by moving downwards, and the liquid can be repeatedly sucked and pushed out by the repeated up and down movement of the piston, so that the uniform mixing of the liquid is realized; at this moment, the purified sample can be configured by controlling a lower pressure valve and a piston through a lower pressure rod, and is transferred to an amplification detection tube for amplification detection;
fluorescence scanning detection: at the specific stage of extension detection, fluorescence scanning motor control fluorescence module is along slide rail horizontal migration, and fluorescence module camera lens passes through the augmentation detecting tube of every card box in proper order, and fluorescence module sends the exciting light simultaneously to gather the fluorescence signal of exciting, and convert the light signal into the signal of telecommunication, send for host computer software and handle.
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