CN111013688B

CN111013688B - qPCR module and modularized qPCR device

Info

Publication number: CN111013688B
Application number: CN201911238626.5A
Authority: CN
Inventors: 朱梦林; 罗成
Original assignee: Shenzhen Gangzhu Medical Technology Co ltd
Current assignee: Shenzhen Gangzhu Medical Technology Co ltd
Priority date: 2019-12-06
Filing date: 2019-12-06
Publication date: 2022-05-17
Anticipated expiration: 2039-12-06
Also published as: CN111013688A

Abstract

The present application relates to a qPCR module and a modular qPCR device, wherein the qPCR module comprises an optical detection module and a thermal cycling module; the optical detection module is provided with a static light source structure and a receiving detection structure matched with the light source structure; the heat circulation module comprises a base and a semiconductor refrigerating piece, the base is arranged in contact with the semiconductor refrigerating piece, and the semiconductor refrigerating piece is used for being arranged in contact with the heat radiating piece; the base is provided with a predetermined number of tube holes and light paths, each tube hole is in one-to-one correspondence with each light path, and the receiving and detecting structure is used for receiving light rays emitted by the light source structure through the light paths. The scheme of the simple and compact modularized qPCR instrument is provided, the static light source and the static heat dissipation are adopted, the internal structure of the instrument is simple, no moving part is needed, the manufacturing is easy, no moving part is needed, and the instrument is stable and reliable; and PCR amplification can be rapidly and stably realized, a modularized design is formed, and PCR instruments with various fluxes can be conveniently formed by combination.

Description

qPCR module and modularized qPCR device

Technical Field

The present application relates to the field of biological detection technology, and in particular, to a qPCR module and a modular qPCR apparatus.

Background

The polymerase chain reaction is a molecular biology technology for amplifying and amplifying specific DNA fragments, which can be regarded as special DNA replication in vitro, and the biggest characteristic of PCR is that trace amount of DNA can be greatly increased. The main process of PCR technology is briefly described as follows. The process of in vitro amplification of DNA is realized by the specific binding of a short piece of single-stranded DNA fragment (also called primer) to a specific region of the template DNA, further using four dNTPs as substrates, and polymerizing the DNA fragment by DNA polymerase along the 3' end of the double-stranded portion formed by the primer and the template. PCR consists of three basic reaction steps of denaturation, annealing and extension. DNA denaturation: at 90-96 deg.c, the double stranded DNA template is broken in the action of heat to form single stranded DNA. DNA annealing: at 25-65 deg.c, the system temperature is lowered, and the primer is combined with the DNA template to form local double strand. DNA extension: at 70-75 deg.C, under the action of Taq enzyme (about 72 deg.C, the activity is optimum), dNTP is used as raw material, and the DNA chain complementary to template is synthesized by extending from 5 'end → 3' end of primer.

qPCR (Real-time Quantitative PCR), namely a Real-time fluorescent Quantitative nucleic acid amplification technology, which is called fluorescent Quantitative PCR (Polymerase chain reaction) for short, is a method for quantitatively analyzing a specific DNA sequence in a sample to be detected by adding a corresponding fluorescent dye or a fluorescent labeled probe on the basis of conventional PCR, detecting the whole PCR process in Real time through fluorescent signal change in the PCR reaction process, and monitoring the total amount of products after each PCR cycle by using fluorescent chemical substances.

The fluorescent quantitative PCR instrument is a reaction instrument for real-time detection, and the functions of the instrument are generally ensured by a thermal cycle system and a fluorescent real-time detection system. The qPCR instrument mainly comprises a heating and refrigerating cycle module, a weak fluorescence detection optical module, a circuit control module, a computer and processing software thereof. Wherein the heating and refrigerating cycle module realizes the temperature cycle of PCR in the forms of air bath, water bath, metal bath and the like; the weak fluorescence detection module consists of a light source part, a light path, a photoelectric sensor and a control part.

However, in the conventional technology, the problem of multi-wavelength fluorescence excitation is generally solved by using a white light source, a turntable and an optical filter, and then fluorescence collection of multiple samples is realized by using a plurality of optical fibers or a mechanical arm. The designed instrument has the advantages of complex structure, difficult realization, more control objects, large and heavy instrument. Moving parts inside the instrument are easily damaged, and multiple optical fibers are easily disordered.

Disclosure of Invention

Based on this, there is a need to provide a qPCR module without moving parts and a modular qPCR apparatus.

A qPCR module comprising an optical detection module and a thermal cycling module;

the optical detection module is provided with a static light source structure and a receiving detection structure matched with the light source structure;

the thermal cycle module comprises a base and a semiconductor refrigerating element, the base is arranged in contact with the semiconductor refrigerating element, and the semiconductor refrigerating element is arranged in contact with the radiating element;

the base is provided with a predetermined number of tube holes and light paths, each tube hole is in one-to-one correspondence with each light path so that each light path is communicated with a corresponding tube hole, and the receiving and detecting structure is used for detecting fluorescence through the light paths.

The qPCR module provides a simple and compact modular qPCR instrument scheme, the implementation mode is compact in structure, a static light source and static heat dissipation are adopted, the internal structure of the instrument is simple, no moving part is needed, the manufacturing is easy, no moving part is needed, and the instrument is stable and reliable; and PCR amplification can be rapidly and stably realized, a modular design is formed, and a PCR instrument with 16, 32, 48 or 96 fluxes can be conveniently formed by combination. And, the instrument is also internally provided with no moving mechanism, so that the instrument can be easily and stably made, the yield of products can be improved, and the transportation is facilitated.

Further, in one embodiment, the hot side of the semiconductor refrigeration element is disposed in contact with the base, and the cold side of the semiconductor refrigeration element is disposed in contact with the heat dissipation element.

In one embodiment, the qPCR module further comprises the heat dissipation element, the semiconductor refrigeration element being located between the base and the heat dissipation element.

In one embodiment, the heat sink is also used for connecting a heat sink.

In one embodiment, the qPCR module further comprises the heat sink, and the heat sink is connected to the heat dissipation member.

In one embodiment, the light source structure is provided with a transparent heating film.

In one embodiment, the receiving and detecting structure has at least one photosensor and/or filter.

In one embodiment, the light source structure has a monochromatic light source.

In one embodiment, the light source structure has a bi-color light source and the optical detection module has a switching circuit connected to the bi-color light source.

In one embodiment, a heat-conducting silicone layer and/or a graphite layer is arranged between the bottom of the base and the semiconductor refrigerating piece; and/or the number of the receiving detection structures is two, the light path comprises two sub light paths, each sub light path is respectively communicated with a corresponding tube hole, and each receiving detection structure detects fluorescence through one sub light path; and/or the base and the heat dissipation piece are both made of metal parts.

A modular qPCR apparatus comprising at least one of any of the qPCR modules.

Drawings

Fig. 1 is a schematic structural diagram according to an embodiment of the present application.

Fig. 2 is another schematic view of the embodiment shown in fig. 1.

FIG. 3 is a schematic sectional view taken along the line A-A of the embodiment shown in FIG. 2.

FIG. 4 is a schematic cross-sectional view along the direction B-B of the embodiment shown in FIG. 2.

Fig. 5 is another schematic view of the embodiment of fig. 2.

FIG. 6 is a schematic cross-sectional view along the direction C-C of the embodiment shown in FIG. 5.

Fig. 7 is another schematic view of the embodiment of fig. 2.

Fig. 8 is another schematic view of the embodiment of fig. 1.

Fig. 9 is another schematic view of the embodiment of fig. 1.

Fig. 10 is another schematic view of the embodiment shown in fig. 1.

Fig. 11 is a schematic structural diagram of another embodiment of the present application.

Detailed Description

In order to make the aforementioned objects, features and advantages of the present application more comprehensible, embodiments accompanying the present application are described in detail below with reference to the accompanying drawings. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present application. This application is capable of embodiments in many different forms than those described herein and that modifications may be made by one skilled in the art without departing from the spirit and scope of the application and it is therefore not intended to be limited to the specific embodiments disclosed below.

It will be understood that when an element is referred to as being "secured to" or "disposed on" another element, it can be directly on the other element or intervening elements may also be present. When an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present. The terms "vertical," "horizontal," "left," "right," and the like as used in the description of the present application are for illustrative purposes only and do not denote a single embodiment.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description of the present application is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. As used in the description of the present application, the term "and/or" includes any and all combinations of one or more of the associated listed items.

In one embodiment of the present application, a qPCR module comprising an optical detection module and a thermal cycling module; the optical detection module is provided with a static light source structure and a receiving detection structure matched with the light source structure; the thermal cycle module comprises a base and a semiconductor refrigerating element, the base is arranged in contact with the semiconductor refrigerating element, and the semiconductor refrigerating element is arranged in contact with the radiating element; the base is provided with a predetermined number of tube holes and light paths, each tube hole is in one-to-one correspondence with each light path so that each light path is communicated with a corresponding tube hole, and the receiving and detecting structure is used for detecting fluorescence through the light paths. The qPCR module provides a simple and compact modular qPCR instrument scheme, the implementation mode is compact in structure, a static light source and static heat dissipation are adopted, the internal structure of the instrument is simple, no moving part is needed, the manufacturing is easy, no moving part is needed, and the instrument is stable and reliable; and PCR amplification can be rapidly and stably realized, a modular design is formed, and a PCR instrument with 16, 32, 48 or 96 fluxes can be conveniently formed by combination. And, the instrument is also internally provided with no moving mechanism, so that the instrument can be easily and stably made, the yield of products can be improved, and the transportation is facilitated.

In one embodiment, a qPCR module comprises some or all of the structure of the following embodiments; that is, the qPCR module includes some or all of the following technical features. In one embodiment, a qPCR module comprises an optical detection module and a thermal cycling module; the optical detection module and the thermal cycle module are core components of qPCR, and the optical detection module detects the PCR process in real time through a fluorescent signal in the PCR amplification process. The thermal cycle module completes thermal cycle of high-temperature denaturation, low-temperature renaturation and proper-temperature extension; in the exponential phase of PCR amplification, the Ct value of the template and the initial copy number of the template have a linear relationship, and therefore, the method can be used as a basis for quantification.

In one embodiment, the optical detection module is provided with a static light source structure and a receiving detection structure matched with the light source structure; in contrast to the conventional technique using a dynamic light source, the static light source structure used in each embodiment is one of the most important inventions of the present application; the light source structure is used for emitting light to the pipe hole carrying the sample to be detected; the receiving detection structure is used for realizing fluorescence detection. In one embodiment, the receiving and detecting structure has at least one photosensor and/or filter. In one embodiment, the light source structure is provided with a transparent heating film. The transparent heating film is used for transmitting light on the one hand and for heating on the other hand. In one embodiment, the light source structure has a monochromatic light source. Alternatively, in one embodiment, the light source structure has a bi-color light source and the optical detection module has a switching circuit connected to the bi-color light source. By the design, the light source structure can flexibly set the light source and the related control mode according to the requirement. The optical detection module is an optical part of the qPCR module, the optical detection module is divided into two independent and static modules by the optical detection module, the light source is a static monochromatic light source, can also be a bicolor light source, can be independently switched by a circuit, and can also be simultaneously lightened by the bicolor light source. The receiving part is also independent and is provided with one or more photoelectric sensors, which can be PD or PMT, and the fluorescence with different wavelengths can be simultaneously received by matching with the optical filter. Further, in one embodiment, the receiving and detecting structure comprises a fluorescence receiver, a predetermined number of receiving optical paths, and a predetermined number of photodetectors; each photoelectric detector is respectively connected with the fluorescence receiver, and each receiving light path is arranged in one-to-one correspondence with each light path, namely each receiving light path corresponds to one light path; and each receiving light path and each photoelectric detector are arranged in one-to-one correspondence and are used for realizing the fluorescence detection of the sample to be detected in the tube hole. The design provides a simple and compact modularized qPCR instrument scheme, the implementation mode is compact in structure, the static light source is adopted, no moving part is needed, the manufacturing is easy, no moving part is needed, and the method is stable and reliable.

In one embodiment, the thermal cycle module comprises a base and a semiconductor refrigeration piece, wherein the base is arranged in contact with the semiconductor refrigeration piece, and the semiconductor refrigeration piece is used for being arranged in contact with a heat dissipation piece; a semiconductor cooler (Thermoelectric cooler) is also one of the important inventions of the present application, and can realize a static cooling effect; the semiconductor refrigeration piece is a device for producing cold by utilizing the thermo-electric effect of a semiconductor, so the semiconductor refrigeration piece is also called as a thermoelectric refrigerator, the semiconductor refrigeration piece is connected with two different metals, and when direct current is switched on, the temperature of one joint is reduced, and the temperature of the other joint is increased; the semiconductor refrigerating element has the characteristics of no noise, no vibration, no need of refrigerant, small volume, light weight and the like, and has the advantages of reliable work, simple and convenient operation and easy cold quantity regulation. Further, in one embodiment, the semiconductor cooling element is also used to achieve a static heating effect. Further, in one embodiment, the hot side of the semiconductor refrigeration element is disposed in contact with the base, and the cold side of the semiconductor refrigeration element is disposed in contact with the heat dissipation element. In one embodiment, the base and the heat sink are both made of metal. In one embodiment, the base is a copper piece or a copper alloy piece, and the heat sink is an aluminum piece or an aluminum alloy piece. By the design, the semiconductor refrigerating sheet is utilized to realize the modularized qPCR module without moving parts, and the qPCR module can be used for a PCR real-time fluorescence detection system. Further, in one embodiment, the semiconductor cooling member is in a sheet shape. In one embodiment, a heat-conducting silicone layer and/or a graphite layer is arranged between the bottom of the base and the semiconductor refrigerating piece. Further, in one embodiment, a heat conductive silicone layer and/or a graphite layer is filled between the bottom of the base and the semiconductor refrigerating element, that is, the heat conductive silicone layer and/or the graphite layer is filled between the bottom of the base and the semiconductor refrigerating element, so as to improve the heat conduction effect. The design provides a simple and compact modularized qPCR instrument scheme, the implementation mode is compact in structure, static heat dissipation is adopted, no moving part is needed, manufacturing is easy, no moving part is needed, and stability and reliability are achieved. And, the instrument is also internally provided with no moving mechanism, so that the instrument can be easily and stably made, the yield of products can be improved, and the transportation is facilitated.

Further, in one embodiment, the base is a solid structure, i.e. the base is a single piece except for the tube hole and the light path; with such a design, the overall heat conduction performance of the base is better. Or, in one embodiment, a cavity is formed in the base, and the cavity is arranged in a non-communication manner with the pipe hole and the light path; the design of the cavity has the advantages of reducing the material consumption of the base, lightening the weight and reducing the cost; disadvantages are reduced and the efficiency of the heat transfer is affected; it is usually necessary to design the base according to the requirement so as to reduce the material consumption of the base while ensuring the control of PCR thermal cycling. Further, in one embodiment, the base is further provided with a temperature sensor hole and a temperature detection position communicated with the temperature sensor hole, and the temperature detection position is used for being communicated with the external environment through the temperature sensor hole; the temperature sensor hole is used for placing a temperature sensor into the base to detect the temperature in the base, and the temperature sensor hole is favorable for accurately controlling the temperature of PCR circulation. In one embodiment, the qPCR module further comprises the temperature sensor. In one embodiment, the temperature sensor is or includes a thermistor, platinum resistor, thermocouple, or the like.

In one embodiment, the base defines a predetermined number of tube holes and light paths, each of the tube holes is disposed in one-to-one correspondence with each of the light paths so that each of the light paths communicates with a corresponding one of the tube holes, and the receiving and detecting structure is configured to detect fluorescence through the light paths. Further, in one embodiment, the tube hole is a square hole or a circular hole, and is used for bearing a square or circular detection container respectively, and a sample to be detected is stored in the detection container; in one embodiment, the tube hole is a through hole or a blind hole; the blind hole is also called a counter bore. In one embodiment, the length of the pipe hole is smaller than that of the detection container, so that the detection container can be conveniently taken and placed. The test receptacle may typically be a cuvette. In one embodiment, the predetermined number is 6 or 8; usually 8 groups can be selected, that is, one qPCR module can perform 8 groups of tests; n qPCR modules are adopted at the same time, so that N x 8 groups of detection can be carried out, and the method is called qPCR module or modularized qPCR detection; moreover, such qPCR modules are advantageous for combining and forming modular qPCR devices as described above, thereby realizing large-scale qPCR detection.

Further, in one embodiment, the number of the receiving and detecting structures is two, the optical path includes two sub optical paths, each sub optical path is respectively communicated with a corresponding one of the tube holes, and each receiving and detecting structure detects fluorescence through one of the sub optical paths. In one embodiment, the receiving and detecting structure includes a fluorescence receiver, a predetermined number of receiving optical paths and a predetermined number of photodetectors, and each receiving optical path is connected to one of the optical paths or one of the sub-optical paths. In one embodiment, the two sub-optical paths of the optical path are located on the same straight line; alternatively, in one embodiment, the two sub-optical paths of the optical path are located on different straight lines, which is advantageous for the two receiving and detecting structures not to interfere with each other. In one embodiment, the lengths of the two sub-optical paths of the optical path are the same or different; in one embodiment, the two sub-paths of the optical path are symmetrically arranged. In one embodiment, a heat-conducting silicone layer and/or a graphite layer is arranged between the bottom of the base and the semiconductor refrigerating piece; the receiving detection structures are two in number, the light path comprises two sub light paths, each sub light path is respectively communicated with one corresponding tube hole, and each receiving detection structure detects fluorescence through one sub light path; the base and the heat dissipation piece are all made of metal. The design is favorable for realizing the fluorescence detection of the sample to be detected in the same pipe hole by adopting two sets of receiving detection structures respectively when in use, and is favorable for realizing the simultaneous detection of double-wavelength and double-fluorescence channels instead of time-sharing detection by matching with the design of the receiving detection structures and the light source structures.

In one embodiment, the qPCR module or thermal cycling module thereof further comprises the heat sink, the semiconductor refrigeration component being located between the base and the heat sink. In one embodiment, the heat sink is also used for connecting a heat sink. Further, in one embodiment, the shape of the heat dissipation element is matched with the shape of the semiconductor refrigeration element; in one embodiment, the semiconductor refrigeration element has a first contact surface, the heat dissipation element has a second contact surface, and the first contact surface and the second contact surface are arranged in a matching manner; in one embodiment, a heat-conducting silicone layer and/or a graphite layer is arranged between the first contact surface and the second contact surface. Further, in one embodiment, the first contact surface and the second contact surface each have a planar shape; in one embodiment, the semiconductor cooling element is sheet-shaped, and the heat dissipation element has a flat surface to contact the semiconductor cooling element. In one embodiment, the heat sink is a copper plate or an aluminum plate. Alternatively, in one embodiment, the first contact surface and the second contact surface have matching tooth shapes to increase the contact area. The contact area is increased, so that the heat conduction effect is improved to a certain extent. The design of the heat dissipation piece is favorable for simplifying the structure of the qPCR module, and in a product with specific application, the modularized qPCR device can be provided with a copper plate on which a plurality of qPCR modules are regularly arranged; or the qPCR module may be self-contained with the heat sink.

In one embodiment, the qPCR module or the thermal cycling module thereof further comprises the heat sink, and the heat sink is connected to the heat dissipation member. Further, in one embodiment, the heat sink is a copper piece, and the heat sink is an aluminum piece, such as an aluminum alloy piece. In one embodiment, a heat-conducting silicone layer and/or a graphite layer is disposed between the heat sink and the heat sink. By the design, on one hand, the copper consumption is saved on the premise of ensuring the heat dissipation performance, so that the cost is saved; on the other hand, various commercially available radiators can be directly adopted, so that the radiator is convenient and direct, and the cost is reduced; on the other hand, the heat dissipation effect of heat conduction is improved.

In each embodiment of this application, the mode that thermal cycle module adopted the semiconductor refrigeration piece is realized, in the product of a specific application, at first be equipped with the base of eight even pipes of an installation standard, the material of base can adopt the metal that the heat transfer effect is good, for example can be materials such as silver, copper, aluminum alloy, the base bottom is with the hot face direct contact of semiconductor refrigeration piece, can paint heat conduction silicone grease, guarantee good heat-conduction and leakproofness, also can middle one deck graphite flake again, reinforcing homogeneity. The cold side of the semiconductor refrigeration piece is a copper sheet or a good heat conductor, and then the semiconductor refrigeration piece is connected to the heat dissipation piece through the copper sheet. According to the design, the thermal cycling module and the optical detection module form an independent qPCR module, if detection modules with 16, 32, 48 or 96 holes are needed, the modules can be combined at will to form required detection flux, each module can be controlled independently or jointly, and therefore the modularized qPCR device is manufactured, multi-channel and high-flux fluorescence static detection can be achieved, PCR amplification can be achieved quickly and stably, modularized design is formed, and the modules can be combined conveniently.

In one embodiment, as shown in fig. 1, only the core module of the qPCR module is shown, and the optical detection module of the qPCR module has a static light source structure 110 and a receiving detection structure matching the light source structure; referring to fig. 2, 3 and 4, the receiving and detecting structure includes a fluorescence receiver 150, a receiving optical path 160 and a photo detector 190(PD detector); referring to fig. 5, 6 and 7, the thermal cycling module of the qPCR module includes a base 120 and a semiconductor cooling element 130, the qPCR module or the thermal cycling module thereof further includes a heat dissipation element 140, the base 120 is disposed in contact with the semiconductor cooling element 130, and the semiconductor cooling element 130 is disposed in contact with the heat dissipation element 140, so that a continuous heat conduction path is formed. Referring to fig. 8, 9 and 10, the base 120 is provided with a predetermined number of tube holes 170 and light paths 200, each of the tube holes 170 and each of the light paths 200 are arranged in a one-to-one correspondence manner so that each of the light paths 200 passes through a corresponding one of the tube holes 170, and the fluorescence receiver 150 of the receiving and detecting structure detects fluorescence of the sample to be detected in the tube hole 170 through the photodetector 190, the receiving light path 160 and the light path 200. The base 120 is further provided with a light path through hole 180 corresponding to the light path 200. In one embodiment, as shown in fig. 1, the light source structure 110 is in a plate shape, i.e. a light source control plate is made; the semiconductor refrigerating piece 130 is in a plate shape, namely, a semiconductor refrigerating piece is manufactured; the heat sink 140 is a copper piece and is shaped like a plate, i.e., a heat conducting copper sheet is made; the orifice 170 is an EP orifice. Further, in one embodiment, as shown in fig. 11, the base 120 further defines a temperature sensor hole 210; in one embodiment, the temperature sensor hole 210 is disposed adjacent to the light path through hole 180. The temperature sensor hole is used for placing a temperature sensor into the base to detect the temperature inside the base, and the temperature of PCR circulation can be accurately controlled. The light source structure comprises a light source control board used for exciting fluorescence, the light source can be a monochromatic LED or a bicolor LED or an LED array and other light sources, a flat heating film and a light filter of the light source are arranged below the light source control board, the light is not shown in the figure, and the light penetrates through the heating film and hits an EP (ultraviolet) tube hole on the base. The receiving optical path is a fluorescence receiving optical path, and the fluorescence receiver is provided with a fluorescence receiving detector control board on which a photoelectric detector, generally a PD, is arranged. The thermal cycle module adopts a semiconductor refrigerating sheet for heating and refrigerating, the hot surface of the semiconductor refrigerating sheet is directly connected with the base through heat-conducting materials with good heat conduction such as heat-conducting silicone grease, the cold surface of the semiconductor refrigerating sheet is connected with the radiating sheet through a heat-conducting copper sheet, and the radiating sheet is not shown in the figure. The base of the thermal cycle module is mainly used for bearing the EP pipe, wherein the base can be solid or hollow, and the EP pipe hole are independent and are not connected into a whole. The semiconductor refrigerating sheet is arranged below the base, the cold surface of the semiconductor refrigerating sheet is connected with the heat conduction copper sheet, and the heat conduction copper sheet is connected with the radiating fin. The top of the base is provided with 8 pipe holes of EP pipes, which can also be called as a hole seat, the two side surfaces are light paths, namely the light paths are provided with two sub light paths, the end part of each sub light path is a light path through hole, and the light paths are mainly communicated with a receiving detection structure to realize the fluorescence detection. The bottom of one side of the hole seat is provided with a mounting hole for mounting a temperature sensor, namely a temperature sensor hole, which is used for mounting a temperature sensor, such as a thermistor, a platinum resistor, a thermocouple and the like, and measuring the temperature of the base so as to control the temperature in the EP pipe. The optical detection plate is mainly a photodetector, mainly a PD, for detecting fluorescence. The design can be independently formed into an 8-hole single-channel or double-channel qPCR instrument, or the qPCR modules can be combined and connected in parallel to form a 16-hole, or 32-hole, or 48-hole, or 96-hole qPCR instrument, and the qPCR instrument can be controlled in a combined manner and independently to form a modular qPCR device, and the qPCR device can realize the simultaneous detection of double-wavelength and double-fluorescence channels without a moving part inside the qPCR device, but not time-sharing detection.

The qPCR module described in the above embodiments can be extended in an array format to increase instrument throughput and detect more samples, and the arrays can be operated either synchronously or separately. In one embodiment, a modular qPCR apparatus comprising at least one qPCR module of any of the embodiments. In one embodiment, the modular qPCR apparatus comprises a plurality of the qPCR modules. In one embodiment, each qPCR module is regularly arranged in an array format. In one embodiment, the qPCR modules operate simultaneously or individually. In one embodiment, a plurality of qPCR modules may be employed to collectively form the modular qPCR apparatus. In one embodiment, the modular qPCR apparatus may further comprise a control apparatus or the like. The design of the modularized qPCR device is beneficial to manufacturing the modularized and multi-flux qPCR instrument in the qPCR field.

Other embodiments of the present application include a qPCR module and a modular qPCR apparatus that can be implemented by combining technical features of the above embodiments.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present application, and the description thereof is more specific and detailed, but not construed as limiting the claims. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the concept of the present application, which falls within the scope of protection of the present application. Therefore, the protection scope of the present application shall be subject to the appended claims.

Claims

1. A qPCR module comprises an optical detection module and a thermal cycling module, and is characterized in that,

the thermal cycle module comprises a base and a semiconductor refrigerating piece, wherein the base is arranged in a solid manner, the base is arranged in contact with the semiconductor refrigerating piece, and the semiconductor refrigerating piece is used for being arranged in contact with a heat radiating piece;

the base is provided with a predetermined number of tube holes and light paths, each tube hole is in one-to-one correspondence with each light path so that each light path is communicated with a corresponding tube hole, and the receiving and detecting structure is used for detecting fluorescence through the light paths;

the receiving detection structures are two in number, the light path comprises two sub light paths, each sub light path is respectively communicated with the corresponding tube hole, and each receiving detection structure detects fluorescence through one sub light path.

2. The qPCR module of claim 1, further comprising the heat sink, the semiconductor refrigeration element being located between the base and the heat sink, a hot side of the semiconductor refrigeration element being disposed in contact with the base, and a cold side of the semiconductor refrigeration element being disposed in contact with the heat sink.

3. The qPCR module according to claim 2, wherein the heat sink is further adapted for connection to a heat sink.

4. The qPCR module of claim 3, further comprising the heat sink disposed in connection with the heat sink.

5. The qPCR module according to claim 1, wherein the light source structure is provided with a transparent heating film.

6. The qPCR module according to claim 1, wherein the receiving detection structure has at least one photosensor and/or filter.

7. The qPCR module according to claim 1, wherein the light source structure has a monochromatic light source.

8. The qPCR module according to claim 1, wherein the light source structure has a bi-color light source and the optical detection module has a switching circuit connected to the bi-color light source.

9. The qPCR module according to any of claims 1 to 8, wherein a thermally conductive silicone layer and/or graphite layer is provided between the bottom of the base and the semiconductor cooling element, the semiconductor cooling element further being configured to achieve a static heating effect;

and/or the base and the heat dissipation piece are both metal parts;

the receiving and detecting structure comprises a fluorescence receiver, a preset number of receiving light paths and a preset number of photoelectric detectors; each photoelectric detector is respectively connected with the fluorescence receiver, and each receiving light path is communicated with one sub light path;

the lengths of the two sub-optical paths of the optical path are the same or different;

the base is further provided with a temperature sensor hole and a temperature detection position communicated with the temperature sensor hole, and the temperature detection position is used for being communicated with the external environment through the temperature sensor hole; the temperature sensor hole is used for placing a temperature sensor into the base, and the qPCR module further comprises the temperature sensor;

the qPCR module or the thermal cycle module thereof also comprises a heat dissipation piece, the heat dissipation piece is a copper plate, and a plurality of qPCR modules are regularly arranged on the copper plate;

the semiconductor refrigeration piece is positioned between the base and the heat dissipation piece; the semiconductor refrigeration piece is provided with a first contact surface, the radiating piece is provided with a second contact surface, the first contact surface and the second contact surface are provided with matched tooth meshing shapes so as to improve the contact area, and a heat conduction silicone grease layer and/or a graphite layer are/is arranged between the first contact surface and the second contact surface.

10. A modular qPCR apparatus comprising at least one qPCR module according to any of claims 1 to 9.