Real-time fluorescence convection PCR instrument

Abstract

The utility model discloses a real-time fluorescent convection PCR (Polymerase Chain Reaction) instrument, relates to the technical field of molecular diagnosis, and solves the technical problem of poor heat dissipation effect of the existing fluorescent quantitative PCR instrument. The device comprises a first host shell, a first upper-layer support and a first lower-layer support are installed in the first host shell, a fluorescence excitation module and a fluorescence detection module are installed on one side of the first upper-layer support, and a mainboard is arranged on the other side of the first upper-layer support. The main board is connected with the first upper-layer support through a multi-layer heat dissipation assembly, a circuit board and a power module are installed on the first lower-layer support, a second host shell is installed on one side of the first host shell, and a convection heating module used for containing a convection PCR reaction tube is installed in the second host shell. According to the utility model, the problems that the circuit module cannot work normally and errors are increased due to overheating can be prevented.

Description

Technical Field

[0001] The utility model relates to the technical field of molecular diagnosis, and more specifically, to a real-time fluorescence convection PCR instrument. Background Art

[0002] The core principle of real-time fluorescence quantitative PCR (Quantitative Real-time Polymerase Chain Reaction) technology is to introduce fluorescent groups during the PCR reaction. The PCR reaction is repeated through high-temperature denaturation, low-temperature annealing and medium-temperature extension steps, so that the target DNA sequence can be efficiently amplified. By real-time monitoring of the intensity changes of the fluorescence signal, the amount of PCR product generated can be accurately calculated, thereby achieving quantitative analysis of the target DNA fragment.

[0003] Traditional real-time fluorescence quantitative PCR instruments need to use semiconductor thermal cyclers in a precise control circuit system to achieve temperature control of the reaction tubes, which usually requires 30-40 cycles and the entire reaction takes 2-3 hours. During this process, a lot of heat will be generated inside the timed fluorescence quantitative PCR instrument, especially the semiconductor thermal cycler part. The heat generated by the semiconductor thermal cycler will cause the temperature inside the real-time fluorescence quantitative PCR instrument to remain at a high state, causing the internal components of the real-time fluorescence quantitative PCR instrument, such as the main board, power supply, fluorescence excitation / detection module, etc. to work in a high temperature environment. In addition, the heat generated by these components themselves will further cause the temperature of the real-time fluorescence quantitative PCR instrument and its components to rise, which is likely to affect the normal operation of the circuit module, and even cause overheating and increase errors. Utility Model Content

[0004] The technical problem to be solved by the utility model is to provide a real-time fluorescence convection PCR instrument in view of the deficiencies of the prior art, so as to solve the technical problem that the existing fluorescence quantitative PCR instrument has poor heat dissipation effect.

[0005] The utility model discloses a real-time fluorescence convection PCR instrument, which comprises a first mainframe shell, in which a first upper bracket and a first lower bracket are installed, a fluorescence excitation module and a fluorescence detection module are installed on one side of the first upper bracket, a mainboard is arranged on the other side of the first upper bracket, and the mainboard is connected to the first upper bracket through a multi-level heat dissipation component, a circuit board and a power supply module are installed on the first lower bracket, a second mainframe shell is installed on one side of the first mainframe shell, a convection heating module for placing a convection PCR reaction tube is installed in the second mainframe shell, an air cooling duct is provided on the side of the second mainframe shell close to the first mainframe shell, and a second heat dissipation component is installed in the second mainframe shell.

[0006] As a further improvement, the multi-level heat dissipation assembly includes a mainboard heat sink, a mainboard upper bracket and a mainboard lower bracket, the mainboard is mounted on the mainboard heat sink, the mainboard heat sink is mounted in the mainboard upper bracket, the mainboard upper bracket is mounted on the mainboard lower bracket, the mainboard heat sink is provided with a plurality of fins that are an integral structure with the mainboard on a side away from the mainboard, and a plurality of heat dissipation circular holes that penetrate all the fins at the same time are provided on both sides of the mainboard heat sink, a first cooling fan is installed on the mainboard lower bracket, an exhaust port connected to the air inlet end of the first cooling fan is provided in the mainboard upper bracket, a cooling fan outlet connected to the air outlet end of the first cooling fan is provided on one side of the mainboard lower bracket, and auxiliary cavities are provided in the mainboard lower brackets on both sides of the first cooling fan, and heat sinks that fit the mainboard upper bracket are installed in the auxiliary cavities.

[0007] Furthermore, the fluorescence excitation module includes an excitation light sleeve, an excitation light source assembly, an excitation light filter and an excitation fiber optic slot, the excitation light source assembly is fixedly installed on one end of the excitation light sleeve, the excitation light filter is installed on the end of the excitation light sleeve close to the excitation light source assembly, the excitation fiber optic slot is installed on the end of the excitation light sleeve away from the excitation light source assembly, an excitation fiber optic through hole is opened in the middle of the excitation fiber optic through hole, the excitation fiber optic, the mirror surface of the excitation light filter, and the emission port of the excitation light source assembly are all located on the same straight line.

[0008] Furthermore, the excitation light source assembly includes an excitation light source LED, an excitation light lens, an excitation light source heat sink and two excitation light bases, the excitation light lens is plugged with a mounting sleeve, the end of the mounting sleeve away from the excitation light lens is plugged with the excitation light source LED, the two excitation light bases are provided with grooves corresponding to the mounting sleeves, the two excitation light bases are fixedly connected, the mounting sleeves are fixedly installed in the holes formed by the two grooves, and the excitation light source LED is fixedly connected to the excitation light source heat sink via thermal conductive glue.

[0009] Furthermore, the fluorescence detection module includes an emitting light sleeve, one end of which is fixedly installed with a fluorescence collection component, an emitting light filter is fixedly installed at one end of the emitting light sleeve close to the fluorescence collection component, and an emitting fiber optic card slot is fixedly installed at one end of the emitting light sleeve away from the fluorescence collection component. A emitting fiber optic through hole is opened in the middle of the emitting fiber optic card slot, and a emitting fiber optic is inserted in the emitting fiber optic through hole. The emitting fiber optic, the mirror surface of the emitting light filter and the receiving port of the fluorescence collection component are all located on the same straight line.

[0010] Furthermore, a second host housing is fixedly connected to one side of the first host housing, and a convection heating module is installed in the second host housing;

[0011] The convection heating module includes an upper constant temperature control unit, an intermediate unit and a lower constant temperature control unit, the upper constant temperature control unit is installed above the intermediate unit, the intermediate unit is installed above the lower constant temperature control unit through a cylindrical gasket, an air insulation layer is provided between the intermediate unit and the lower constant temperature control unit, and a plurality of convection heating chambers are provided between the upper constant temperature control unit, the intermediate unit and the lower constant temperature control unit, and the convection heating chambers are connected to the air insulation layer.

[0012] Furthermore, the upper constant temperature control unit is provided with a plurality of first reaction holes for placing convection PCR reaction tubes, the middle unit is provided with a plurality of second reaction holes corresponding one-to-one to the first reaction holes, and the lower constant temperature control unit is provided with a plurality of third reaction holes corresponding one-to-one to the second reaction holes, the aperture of the second reaction hole is larger than the aperture of the first reaction hole and the third reaction hole, and the first reaction hole, the second reaction hole and the third reaction hole constitute a stepped convection heating chamber with a large middle part and small ends.

[0013] Furthermore, a detection optical fiber slot is fixedly installed in the air insulation layer, and a convection PCR reaction tube through hole corresponding to the third reaction hole is provided on the detection optical fiber slot, a first optical fiber through hole is provided at one end of the detection optical fiber slot, and a second optical fiber through hole is provided at the other end of the detection optical fiber slot, one end of the excitation optical fiber is plugged into the first optical fiber through hole, and one end of the emission optical fiber is plugged into the second optical fiber through hole.

[0014] Furthermore, a cooling fan is fixedly installed on one side of the second host housing.

[0015] Furthermore, a touch screen is installed on the first host shell, the touch screen is electrically connected to the mainboard, and a heat dissipation groove is opened in the middle of the upper bracket of the mainboard.

[0016] Beneficial Effects

[0017] The advantages of the utility model are:

[0018] The utility model realizes PCR detection of samples by arranging a fluorescence excitation module, a fluorescence detection module, a circuit board, a power module and a convection heating module; the utility model arranges a multi-level heat dissipation component and a second heat dissipation component, and further dissipates heat to the mainboard through the heat sink, the first heat dissipation fan and the heat dissipation pipe in the multi-level heat dissipation component, and the second heat dissipation component dissipates heat inside the second host shell to prevent the problem of circuit module not being able to work normally and error increase due to overheating; the utility model ensures that the convection heating component in the second host shell has a stable temperature environment by assembling the first host shell and the second host shell separately, and at the same time can effectively prevent the convection heating component from having an adverse effect on the circuit elements in the first host shell. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 It is a front schematic diagram of the convection PCR instrument of the present invention;

[0020] Figure 2 It is a schematic diagram of the back side of the convection PCR instrument of the present invention;

[0021] Figure 3 This is a diagram of the internal structure of the second mainframe housing of the utility model;

[0022] Figure 4 It is a structural schematic diagram of the first upper support and the first lower support of the utility model;

[0023] Figure 5 It is a disassembled schematic diagram of the multi-level heat dissipation component of the utility model;

[0024] Figure 6 It is a structural schematic diagram of the fluorescence excitation module in the utility model;

[0025] Figure 7 It is a structural schematic diagram of the fluorescence collection module in the utility model;

[0026] Figure 8 It is a structural schematic diagram of the excitation light source assembly in the utility model;

[0027] Fig. 9 This is a schematic diagram of the overall structure of the convection heating assembly of the utility model;

[0028] Fig.10 This is a disassembled diagram of the convection heating assembly of the utility model;

[0029] Fig.11 This is a schematic diagram of the structure of the convection PCR reaction tube in the utility model;

[0030] Fig.12 This is a schematic diagram of the structure of the optical fiber card slot in the detection of the utility model;

[0031] Fig.13 This is a diagram of the coordination of the detection optical fiber, the detection optical fiber slot, the convection heating assembly and the convection PCR reaction tube in the utility model;

[0032] Fig.14 It is a real-time fluorescence convection PCR amplification curve diagram of the utility model for different enterohemorrhagic Escherichia coli O157:H7 DNA concentrations;

[0033] Fig.15 This is the gel electrophoresis diagram of the real-time fluorescence convection PCR amplification product of different enterohemorrhagic Escherichia coli O157:H7 DNA concentrations of the utility model.

[0034] in, Fig.15 The experimental conditions for lanes 1 to 4 are that the length of the amplified DNA fragment is 168 bp, and the DNA concentrations are 2.85 ng / μL (lane 1), 2.85×10-2 ng / μL (lane 2), 2.85×10-4 ng / μL (lane 3), and 2.85×10-6 ng / μL (lane 4);

[0035] Fig.16 It is a real-time fluorescence convection PCR amplification curve diagram of the utility model under the same enterohemorrhagic Escherichia coli O157:H7 DNA concentration;

[0036] Fig.17 This is the gel electrophoresis diagram of the real-time fluorescence convection PCR amplification product of the utility model at the same enterohemorrhagic Escherichia coli O157:H7 DNA concentration.

[0037] in, Fig.17 The experimental conditions for lanes 1 to 5 are that the length of the amplified DNA fragment is 168 bp and the DNA concentration is 2.85×10-2 ng / μL.

[0038] Wherein: 1-first host shell, 2-mainboard, 3-mainboard radiator, 4-mainboard upper bracket, 5-mainboard lower bracket, 6-cooling fan outlet, 7-air cooling duct, 8-heat dissipation circular hole, 9-auxiliary cavity, 10-first upper bracket, 11-first lower bracket, 12-fluorescence excitation module, 121-excitation light sleeve, 122-excitation light source assembly, 123-excitation light filter, 124-excitation fiber card slot, 125-excitation fiber through hole, 126-excitation fiber, 127-excitation light source LED, 129-excitation light lens, 1210-installation sleeve, 1211-excitation light base, 1212-excitation light source heat sink, 13-fluorescence detection module, 131-emission light sleeve, 132-fluorescence collection assembly, 133-emission light filter, 134-emission Fiber optic slot, 135-fiber optic through hole, 136-fiber optic, 14-circuit board, 15-power module, 16-second host housing, 17-convection heating module, 171-upper constant temperature control unit, 172-middle unit, 173-lower constant temperature control unit, 174-cylindrical gasket, 175-air insulation layer, 176-convection heating chamber, 177-first reaction hole, 178-second reaction hole, 179-third reaction hole, 1710-detection fiber optic slot, 1711-convection PCR reaction tube through hole, 1712-first fiber optic through hole, 1713-second fiber optic through hole, 18-touch screen, 19-convection PCR reaction tube, 20-heat sink, 21-USB expansion port, 22-power switch, 23-first cooling fan, 24-second cooling fan. DETAILED DESCRIPTION

[0039] The utility model is further described below in conjunction with the embodiments, but does not constitute any limitation to the utility model. Any limited modifications made by anyone within the scope of the claims of the utility model are still within the scope of the claims of the utility model.

[0040] See also Figure 1-Figure 17 ,like Figure 4 As shown, a real-time fluorescence convection PCR instrument of the utility model comprises a first mainframe housing 1, a first upper support 10 and a first lower support 11 are installed in the first mainframe housing 1, a fluorescence excitation module 12 and a fluorescence detection module 13 are installed on one side of the first upper support 10, and a mainboard 2 is provided on the other side of the first upper support 10, and the model of the mainboard 2 is NanoPiM4B. The mainboard 2 is connected to the first upper support 10 through a multi-level heat dissipation component, a circuit board 14 and a power module 15 are installed on the first lower support 11, a second mainframe housing 16 is installed on one side of the first mainframe housing 1, a convection heating module 17 for placing a convection PCR reaction tube 19 is installed in the second mainframe housing 16, an air cooling duct 7 is provided on the side of the second mainframe housing 16 close to the first mainframe housing 1, and a second heat dissipation component is installed in the second mainframe housing 16.

[0041] like Figure 3 As shown, the second heat dissipation component is a second heat dissipation fan 24, which is used to dissipate heat inside the second host housing 16.

[0042] like Figure 5 As shown, the multi-level heat dissipation component includes a mainboard heat sink 3, a mainboard upper bracket 4 and a mainboard lower bracket 5. The mainboard 2 is mounted on the mainboard heat sink 3, the mainboard heat sink 3 is mounted in the mainboard upper bracket 4, the mainboard upper bracket 4 is mounted on the mainboard lower bracket 5, a side of the mainboard heat sink 3 away from the mainboard 2 is provided with a plurality of fins which are an integral structure with the mainboard heat sink 3, both sides of the mainboard heat sink 3 are provided with a plurality of heat dissipation circular holes 8 which simultaneously penetrate all the fins, a first heat dissipation fan 23 is installed on the mainboard lower bracket 5, an air vent connected to the air inlet end of the first heat dissipation fan 23 is opened in the mainboard upper bracket 4, a heat dissipation fan air outlet 6 connected to the air outlet end of the first heat dissipation fan 23 is provided on one side of the mainboard lower bracket 5, auxiliary cavities 9 are provided in the mainboard lower bracket 5 on both sides of the first heat dissipation fan 23, and heat dissipation fins which fit the mainboard upper bracket 4 are installed in the auxiliary cavities 9. The heat dissipation fins, the heat dissipation circular holes 8 and the first heat dissipation fan 23 are all used to dissipate heat from the mainboard 2. The first cooling fan 23 is started to discharge the heat generated by the mainboard 2 from the cooling fan outlet 6 through the first cooling fan 23 .

[0043] like Figure 6As shown, the fluorescence excitation module 12 includes an excitation light sleeve 121, an excitation light source assembly 122, an excitation light filter 123, and an excitation fiber optic slot 124. The excitation light source assembly 122 is fixedly installed at one end of the excitation light sleeve 121, and the excitation light filter 123 is installed at the end of the excitation light sleeve 121 close to the excitation light source assembly 122. The excitation light filter 123 is a high-pass filter with a central wavelength of 490±10nm. An excitation fiber optic slot 124 is installed at the end of the excitation light sleeve 121 away from the excitation light source assembly 122. An excitation fiber optic through hole 125 is opened in the middle of the excitation fiber optic slot 124. An excitation fiber optic 126 is inserted into the excitation fiber optic through hole 125. The excitation fiber optic 126 is a high-temperature resistant fiber of Boyijingke. The excitation fiber optic 126, the mirror surface of the excitation light filter 123, and the emission port of the excitation light source assembly 122 are all located on the same straight line. The excitation light emitted by the excitation light source assembly 122 passes through the excitation light filter 123 and the excitation optical fiber 126 in sequence and irradiates the reaction solution in the convection PCR reaction tube 19 .

[0044] like Figure 8 As shown, the excitation light source assembly 122 includes an excitation light source LED 127, an excitation light lens 129, an excitation light source heat sink 1212 and two excitation light bases 1211. The excitation light source LED 127 is a CREE packaged LED with a central wavelength of 485±5nm, the excitation light lens 129 is an optical grade PMMA lens, and the excitation light source heat sink 1212 is made of brass.

[0045] The excitation light lens 129 is plugged with a mounting sleeve 1210, and the end of the mounting sleeve 1210 away from the excitation light lens 123 is plugged with the excitation light source LED 127. The two excitation light bases 1211 are provided with grooves corresponding to the mounting sleeve 1210. The two excitation light bases 1211 are fixedly connected, and the mounting sleeve 1210 is fixedly installed in the hole formed by the two grooves. The excitation light source LED 127 is fixedly connected to the excitation light source heat sink 1212 via thermal conductive glue (thermal conductive silicone).

[0046] like Figure 7 As shown, the fluorescence detection module 13 includes an emission light sleeve 131, and a fluorescence collection component 132 is fixedly installed at one end of the emission light sleeve 131, and the fluorescence collection component 132 is a Sony IMX291 high-performance CMOS camera. An emission light filter 133 is fixedly installed at one end of the emission light sleeve 131 close to the fluorescence collection component 132, and the emission light filter 133 is a narrow bandwidth filter with a central wavelength of 522±10nm. An emission fiber optic card slot 134 is fixedly installed at one end of the emission light sleeve 131 away from the fluorescence collection component 132, and an emission fiber optic through hole 135 is opened in the middle of the emission fiber optic card slot 134, and an emission fiber optic through hole 135 is inserted into the emission fiber optic through hole 135. The emission fiber optic 136 is a high-temperature resistant fiber of Boyijingke.

[0047] The emission optical fiber 136, the mirror surface of the emission light filter 133 and the receiving port of the fluorescence collection component 132 are all located on the same straight line. The emission light emitted by the reaction solution in the convection PCR reaction tube 19 passes through the emission optical fiber 136 and the emission light filter 133 in sequence to reach the fluorescence collection component 132.

[0048] like Fig. 9 As shown, the convection heating module 17 includes an upper constant temperature control unit 171, an intermediate unit 172 and a lower constant temperature control unit 173, the upper constant temperature control unit 171 is installed above the intermediate unit 172, the intermediate unit 172 is installed above the lower constant temperature control unit 173 through a cylindrical gasket 174, an air insulation layer 175 is provided between the intermediate unit 172 and the lower constant temperature control unit 173, a plurality of convection heating chambers 176 are provided between the upper constant temperature control unit 171, the intermediate unit 172 and the lower constant temperature control unit 173, and the convection heating chambers 176 are connected to the air insulation layer 175.

[0049] like Fig.10 As shown, the upper constant temperature control unit 171 is provided with a plurality of first reaction holes 177 for placing convection PCR reaction tubes 19, the middle unit 172 is provided with a plurality of second reaction holes 178 corresponding one-to-one to the first reaction holes 177, and the lower constant temperature control unit 173 is provided with a plurality of third reaction holes 179 corresponding one-to-one to the second reaction holes 178, the aperture of the second reaction hole 178 is larger than the aperture of the first reaction hole 177 and the third reaction hole 179, and the first reaction hole 177, the second reaction hole 178 and the third reaction hole 179 constitute a stepped convection heating chamber 176 with a large middle portion and small ends.

[0050] The upper constant temperature control unit 171, the middle unit 172 and the lower constant temperature control unit 173 are all provided with a temperature acquisition sensor element interface and a heating element interface, the temperature acquisition sensor element is installed in the temperature acquisition sensor element interface, the heating element is installed in the heating element interface, and the plurality of temperature acquisition sensor elements and heating elements are electrically connected to the circuit board 14. The circuit board 14 is used to control the temperature acquisition sensor element and the heating element.

[0051] like Fig.12 As shown, a detection optical fiber slot 1710 is fixedly installed in the air insulation layer 175, and a convection PCR reaction tube through hole 1711 corresponding to the third reaction hole 179 is provided on the detection optical fiber slot 1710, a first optical fiber through hole 1712 is provided at one end of the detection optical fiber slot 1710, and a second optical fiber through hole 1713 is opened at the other end of the detection optical fiber slot 1710, one end of the excitation optical fiber 126 is inserted into the first optical fiber through hole 1712, and one end of the emission optical fiber 136 is inserted into the second optical fiber through hole 1713.

[0052] A second cooling fan 24 is fixedly mounted on one side of the second host housing 16. The second cooling fan 24 is electrically connected to the power module 15. The first cooling fan 23 and the second cooling fan 24 are both 24V silent centrifugal fans. The central axis of the first cooling fan 23 and the central axis of the convection heating module 17 are located on the same straight line, and the distance between the two is 70 mm.

[0053] A touch screen 18 is installed on the first host housing 1, and the touch screen 18 is electrically connected to the mainboard 2. The touch screen 18 is a capacitive IPS touch screen, which is convenient for operation and viewing of test results.

[0054] The back of the host housing 1 is provided with a power switch 22, a heat sink 20 and a USB expansion port 21 (USB3.0 interface). The power switch 22 is electrically connected to the power module 15. The power switch 22 is used to control the power module 5 to transmit power to the power-consuming device. The heat sink 20 faces the heat dissipation fan outlet 6 and is used to dissipate heat inside the first host housing 1. The USB expansion port is connected to the motherboard 2 for reading data.

[0055] For the above-mentioned real-time fluorescence convection PCR instrument, this paper illustrates the technical effect of using a multi-level heat dissipation component and an air cooling duct 7 on the experiment through the following two application methods:

[0056] Application method 1: Using SYBR Green I as a fluorescent dye, real-time fluorescence convection PCR technology is used to detect enterohemorrhagic Escherichia coli O157:H7 as an example. The process of the example is as follows:

[0057] (1) Take 0.4 μL of the extracted E. coli DNA and 2X

[0058] SGExcelFastSYBRMixture 10μL, upstream primer, downstream primer 0.4μL each, RNase-Free ddH2O 8.8μL, to prepare 20μL reaction solution.

[0059] (2) Use a pipette to drop the prepared reaction solution into the convection PCR reaction tube 19, and centrifuge to evenly mix the reaction solution.

[0060] (3) Use a pipette to further add 1 μL of dimethyl silicone oil into the convection PCR reaction tube 19, and seal the reaction solution to prevent the solution from evaporating.

[0061] (4) Insert the convection PCR reaction tube 19 filled with the reaction solution into the convection heating module 17 in the second main body housing 16 .

[0062] (5) On the touch screen 18, the temperature of the upper constant temperature control unit 171 is set to 98°C, the temperature of the lower constant temperature control unit 173 is set to 53°C, the response time is set to 30 min, the LED drive current is set to 60 mA, and the parameters of the fluorescence collection component 132 are set to camera exposure value 5000, gain 0, brightness 30, and white balance 6400K.

[0063] (6) Click the start button on the touch screen 18 to start the detection process; start the heating module, control the temperature at the set temperature, automatically start the excitation light source LED every 15 seconds and collect a frame of image, use the multi-threaded algorithm to analyze the average gray value of the detection area and store it in the data file, and display the amplification curve in real time in the human-computer interaction module. Start the first cooling fan 23 and the second cooling fan 24 to dissipate heat.

[0064] (7) When the reaction process is completed, the instrument automatically turns off the heating, stores the experimental data in a txt file in the computer, and turns off the first cooling fan 23 and the second cooling fan 2.

[0065] (8) Further data processing is performed to obtain the required real-time fluorescence amplification curve.

[0066] (9) Perform gel electrophoresis analysis on the amplified products of real-time fluorescence convection PCR to obtain the required gel electrophoresis pattern.

[0067] The experimental results are as follows Fig.14 and Fig.15 As shown, it can be seen from the figure that the real-time fluorescence convection PCR instrument of the utility model can effectively perform real-time amplification and detection on DNA samples of different concentrations, and has good detection sensitivity and specificity.

[0068] Application method 2: Using SYBR GreenⅠ as the fluorescent dye, real-time fluorescence convection PCR technology was used to detect enterohemorrhagic Escherichia coli O157:H7 as an example. The operation process was the same as application method 1. The DNA concentration in the reaction solution was 2.85×10-3ng / μL, and the test was repeated 5 times.

[0069] The experimental results are as follows Fig.16 and Fig.17 As shown in the figure, it can be seen that the instrument can achieve effective amplification in repeated experiments under low and medium DNA target concentration conditions, and can clearly distinguish the positive sample group from the negative control group (background). Each amplification curve shows a similar trend, which proves that the instrument has good stability.

[0070] The above is only a preferred embodiment of the present invention. It should be pointed out that for those skilled in the art, several modifications and improvements can be made without departing from the structure of the present invention, which will not affect the effect of the implementation of the present invention and the practicality of the patent.

Claims

1. A real-time fluorescence convection PCR instrument, characterized in that: The invention comprises a first mainframe housing (1), wherein a first upper support (10) and a first lower support (11) are installed in the first mainframe housing (1), a fluorescence excitation module (12) and a fluorescence detection module (13) are installed on one side of the first upper support (10), a mainboard (2) is provided on the other side of the first upper support (10), and the mainboard (2) is connected to the first upper support (10) through a multi-layer heat dissipation component, a circuit board (14) and a power module (15) are installed on the first lower support (11), a second mainframe housing (16) is installed on one side of the first mainframe housing (1), a convection heating module (17) for placing a convection PCR reaction tube (19) is installed in the second mainframe housing (16), an air cooling duct (7) is provided on the side of the second mainframe housing (16) close to the first mainframe housing (1), and a second heat dissipation component is installed in the second mainframe housing (16).

2. A real-time fluorescence convection PCR instrument according to claim 1, characterized in that: The multi-layer heat dissipation component comprises a mainboard heat sink (3), a mainboard upper bracket (4) and a mainboard lower bracket (5); the mainboard (2) is mounted on the mainboard heat sink (3), the mainboard heat sink (3) is mounted in the mainboard upper bracket (4), the mainboard upper bracket (4) is mounted on the mainboard lower bracket (5); a side of the mainboard heat sink (3) away from the mainboard (2) is provided with a plurality of fins which are integrally formed with the mainboard heat sink (3), and both sides of the mainboard heat sink (3) are provided with a plurality of heat dissipation holes which penetrate all the fins at the same time. The mainboard lower bracket (5) is provided with a first cooling fan (23); the mainboard upper bracket (4) is provided with an exhaust port connected to the air inlet of the first cooling fan (23); one side of the mainboard lower bracket (5) is provided with a cooling fan air outlet (6) connected to the air outlet of the first cooling fan (23); auxiliary cavities (9) are provided in the mainboard lower bracket (5) on both sides of the first cooling fan (23); and heat sinks that fit the mainboard upper bracket (4) are installed in the auxiliary cavities (9).

3. A real-time fluorescence convection PCR instrument according to claim 1, characterized in that: The fluorescence excitation module (12) comprises an excitation light sleeve (121), an excitation light source assembly (122), an excitation light filter (123) and an excitation optical fiber slot (124); the excitation light source assembly (122) is fixedly mounted on one end of the excitation light sleeve (121); the excitation light filter (123) is mounted on the end of the excitation light sleeve (121) close to the excitation light source assembly (122); the excitation optical fiber slot (124) is mounted on the end of the excitation light sleeve (121) away from the excitation light source assembly (122); an excitation optical fiber through hole (125) is provided in the middle of the excitation optical fiber slot (124); an excitation optical fiber (126) is inserted into the excitation optical fiber through hole (125); the excitation optical fiber (126), the mirror surface of the excitation light filter (123) and the emission port of the excitation light source assembly (122) are all located on the same straight line.

4. A real-time fluorescence convection PCR instrument according to claim 3, characterized in that: The excitation light source assembly (122) comprises an excitation light source LED (127), an excitation light lens (129), an excitation light source heat sink (1212) and two excitation light bases (1211); the excitation light lens (129) is plugged with a mounting sleeve (1210); the excitation light source LED (127) is plugged into one end of the mounting sleeve (1210) away from the excitation light lens (129); the two excitation light bases (1211) are provided with grooves corresponding to the mounting sleeves (1210); the two excitation light bases (1211) are fixedly connected; the mounting sleeves (1210) are fixedly installed in a hole formed by the two grooves; and the excitation light source LED (127) is fixedly connected to the excitation light source heat sink (1212) via a thermally conductive adhesive.

5. A real-time fluorescence convection PCR instrument according to claim 3, characterized in that: The fluorescence detection module (13) comprises an emission light sleeve (131), one end of the emission light sleeve (131) is fixedly mounted with a fluorescence collection component (132), an emission light filter (133) is fixedly mounted inside the emission light sleeve (131) at one end close to the fluorescence collection component (132), an emission optical fiber card slot (134) is fixedly mounted inside the emission light sleeve (131) at one end away from the fluorescence collection component (132), a emission optical fiber through hole (135) is provided in the middle of the emission optical fiber card slot (134), an emission optical fiber (136) is inserted into the emission optical fiber through hole (135), and the emission optical fiber (136), the mirror surface of the emission light filter (133) and the receiving port of the fluorescence collection component (132) are all located on the same straight line.

6. A real-time fluorescence convection PCR instrument according to claim 5, characterized in that: The convection heating module (17) comprises an upper constant temperature control unit (171), an intermediate unit (172) and a lower constant temperature control unit (173); the upper constant temperature control unit (171) is installed above the intermediate unit (172); the intermediate unit (172) is installed above the lower constant temperature control unit (173) via a cylindrical gasket (174); an air insulation layer (175) is provided between the intermediate unit (172) and the lower constant temperature control unit (173); a plurality of convection heating chambers (176) are provided between the upper constant temperature control unit (171), the intermediate unit (172) and the lower constant temperature control unit (173); and the convection heating chambers (176) are connected to the air insulation layer (175).

7. A real-time fluorescence convection PCR instrument according to claim 6, characterized in that: The upper constant temperature control unit (171) is provided with a plurality of first reaction holes (177) for placing convection PCR reaction tubes (19); the middle unit (172) is provided with a plurality of second reaction holes (178) corresponding one-to-one to the first reaction holes (177); the lower constant temperature control unit (173) is provided with a plurality of third reaction holes (179) corresponding one-to-one to the second reaction holes (178); the aperture of the second reaction hole (178) is larger than the aperture of the first reaction hole (177) and the third reaction hole (179); the first reaction hole (177), the second reaction hole (178) and the third reaction hole (179) constitute a stepped convection heating chamber (176) with a large middle portion and small ends.

8. A real-time fluorescence convection PCR instrument according to claim 7, characterized in that: A detection optical fiber slot (1710) is fixedly installed in the air insulation layer (175); a convection PCR reaction tube through hole (1711) corresponding to the third reaction hole (179) is provided on the detection optical fiber slot (1710); a first optical fiber through hole (1712) is provided at one end of the detection optical fiber slot (1710); a second optical fiber through hole (1713) is provided at the other end of the detection optical fiber slot (1710); one end of the excitation optical fiber (126) is plugged into the first optical fiber through hole (1712) through an air cooling duct (7); and one end of the emission optical fiber (136) is plugged into the second optical fiber through hole (1713) through an air cooling duct (7).

9. A real-time fluorescence convection PCR instrument according to claim 1, characterized in that: The second heat dissipation component is a second heat dissipation fan (24).

10. A real-time fluorescence convection PCR instrument according to claim 2, characterized in that: A touch screen (18) is installed on the first host housing (1), the touch screen (18) is electrically connected to the mainboard (2), and a heat dissipation groove (20) is provided in the middle of the mainboard upper bracket (4).

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