Carbapenem drug resistance gene detection device

Abstract

The invention discloses a carbapenem drug resistance gene detection device, which comprises a detection box, a flip plate, a partition plate, a sample plate, a fluorescence detection unit, a thermal cycle unit and a computer controller, the corner end of the flip plate is rotatably installed on the top of the detection box, the partition plate is fixedly installed on the inner wall of the detection box, and the thermal cycle unit is fixedly installed on the bottom of the detection box. And the mounting column of the sample plate is fixedly mounted on the interlayer plate. The temperature in the sample holes of the sample plate is increased through high-speed flowing silicone oil, the denaturation temperature box, the annealing temperature box and the extension temperature box correspond to the temperature change required by the denaturation, annealing and extension detection steps, and the cold force temperature box is used for regulating the temperature balance of the silicone oil in the denaturation temperature box; and the denaturation temperature is quickly transited to the annealing temperature, the temperature of the sample hole is quickly reduced, and the high-speed flowing silicone oil can quickly convert the denaturation, annealing and extension corresponding temperature, so that the reaction efficiency is improved, non-specific products are reduced, the data quality is improved, and the method is suitable for high-throughput detection and the like.

Description

Technical Field

[0001] The present invention relates to the technical field of drug resistance gene detection, and in particular to a carbapenem resistance gene detection device. Background Art

[0002] Carbapenems are a class of broad-spectrum β-lactam antibiotics that are widely used to treat multidrug-resistant bacterial infections. However, with the widespread use of carbapenems, bacterial resistance to these antibiotics has gradually increased, making the detection of carbapenem resistance genes particularly important. Although existing carbapenem resistance gene detection devices have met laboratory and clinical needs to a certain extent, they still have many shortcomings in practical applications, especially in the design of temperature control and heating units.

[0003] Existing carbapenem resistance gene detection devices usually use quantitative polymerase chain reaction (qPCR) technology, which realizes quantitative analysis of target genes by real-time monitoring of changes in fluorescence signals. In the qPCR process, denaturation, annealing and extension are three key temperature control steps. However, existing devices have obvious problems in these temperature conversion processes. The temperature conversion time of denaturation, annealing and extension is usually around 4 seconds. Although such conversion time can meet basic experimental needs, it is not flexible enough in actual operation. Fast and accurate temperature conversion is crucial to improving the efficiency and specificity of PCR reactions, and existing devices perform poorly in this regard.

[0004] A longer temperature conversion time will increase the overall time of the PCR reaction. Each cycle of denaturation, annealing, and extension steps requires a certain amount of time. If the temperature conversion time is long, the entire PCR process will become lengthy, reducing the detection efficiency. This is particularly evident in high-throughput detection, where long temperature conversions will seriously affect the throughput and efficiency of the detection. In addition, a longer temperature conversion time may also lead to an increase in non-specific products. During the process from denaturation to annealing, if the temperature drops too slowly, the primers have more time to bind to non-complementary sequences, thereby increasing the possibility of non-specific amplification. Similarly, during the process from annealing to extension, if the temperature rises too slowly, the DNA polymerase has more time to synthesize DNA on non-specifically bound primers, further increasing the generation of non-specific products.

[0005] The heating unit of the existing carbapenem resistance gene detection device is usually electric heating. Although this heating method can achieve rapid temperature rise, it has some problems in practical application. Electric heating elements are prone to cracks, breakage or other damage after long-term use, which not only affects the accuracy of temperature control, but also may lead to equipment failure and increased maintenance costs. In addition, the heating and cooling speeds of electric heating elements are limited, and rapid and uniform temperature changes cannot be achieved, which to a certain extent limits the efficiency and specificity of PCR reactions. Especially in high-throughput detection, frequent temperature conversions will place a heavy burden on the heating elements, accelerating their aging and damage.

[0006] Existing carbapenem resistance gene detection devices also lack flexibility in temperature control. Inflexible temperature conversion means that the device has poor adaptability when dealing with different types of samples and different experimental conditions. This not only affects the accuracy and reliability of the experiment, but also increases the complexity and difficulty of experimental operations.

[0007] Therefore, how to provide a carbapenem resistance gene detection device is a problem that those skilled in the art urgently need to solve. Summary of the invention

[0008] One object of the present invention is to provide a carbapenem resistance gene detection device. The present invention heats the temperature in the sample well of the sample plate by high-speed flowing silicone oil, and then uses a denaturation temperature box, an annealing temperature box and an extension temperature box to adjust the temperature changes required for the denaturation, annealing and extension detection steps. The cold force temperature box is used to regulate the temperature balance of the silicone oil in the denaturation temperature box, and the denaturation temperature quickly transitions to the annealing temperature, and the temperature in the sample well is quickly reduced. The high-speed flowing silicone oil can quickly convert the corresponding temperatures of denaturation, annealing and extension, thereby improving reaction efficiency, reducing non-specific products, improving data quality and adapting to high-throughput detection.

[0009] A carbapenem resistance gene detection device according to an embodiment of the present invention comprises a detection box, a flap plate, an interlayer plate, a sample plate, a fluorescence detection unit, a thermal cycle unit and a computer controller, wherein the corner end of the flap plate is rotatably mounted on the top of the detection box, the interlayer plate is fixedly mounted on the inner wall of the detection box, the mounting column of the sample plate is fixedly mounted on the interlayer plate, the fluorescence detection unit is slidably mounted inside the detection box, the thermal cycle unit is fixedly mounted on the inner bottom of the detection box, and the computer controller is fixedly mounted on the inner bottom of the detection box; A sample hole is provided on the top of the sample plate, and a plurality of the sample holes are provided, and the plurality of the sample holes are arranged in a matrix; The fluorescence detection unit includes a displacement assembly, a displacement sensor and a fluorescence detector, wherein the displacement assembly is mounted on the inner top of the detection box, the displacement sensor is fixedly mounted on the outer wall of the fluorescence detector, and the fluorescence detector is threadedly mounted on the displacement assembly; The thermal cycle unit includes a heat supply component, an airtight component and a first temperature sensor, wherein the bottom of the heat supply component is fixedly mounted on the inner bottom of the detection box, the airtight component is fixedly mounted on the inner bottom of the detection box, the first temperature sensor is fixedly mounted on the bottom of the sample plate, and a plurality of first temperature sensors are provided, the first temperature sensors match the number of sample holes, each of the first temperature sensors corresponds to a sample hole, and the first temperature sensor is located directly below the bottom of the sample hole; The heat supply component includes a spiral channel, which is opened inside the sample plate. There are multiple spiral channels, and the number of the spiral channels matches the number of sample holes. Each spiral channel is sleeved on a corresponding sample hole. The airtight component includes an airtight rubber ring, which is fixedly installed on the inner top of the sample hole. There are multiple airtight rubber rings, and the number of the airtight rubber rings matches the number of sample holes. Each of the airtight rubber rings corresponds to a sample hole.

[0010] Furthermore, a first signal receiving port is provided on an outer wall of the detection box, and a second signal receiving port is provided on an outer wall of the detection box.

[0011] Further, the displacement assembly includes a displacement motor, a threaded rod and a guide rod, wherein the displacement motor is fixedly mounted on the inner wall of the detection box, both ends of the threaded rod are rotatably mounted on the inner top of the detection box, the threaded rod is fixedly mounted on the rotating shaft of the displacement motor, the guide rod is fixedly mounted on the inner top of the detection box, and the guide rod is located directly above the threaded rod; The displacement assembly also includes a threaded sleeve and a guide slide cylinder. The threaded sleeve is threadedly mounted on the threaded rod. The threaded sleeve is fixedly mounted on both ends of the fluorescence detector. The guide slide cylinder is slidably mounted on the guide rod. The guide slide cylinder is fixedly mounted on both ends of the fluorescence detector.

[0012] Furthermore, the heat supply component also includes a denaturation temperature box, an annealing temperature box, an extension temperature box, a cold force temperature box, a second temperature sensor and a heating tube, wherein the bottom of the denaturation temperature box is fixedly installed on the inner bottom of the detection box, the bottom of the annealing temperature box is fixedly installed on the inner bottom of the detection box, the bottom of the extension temperature box is fixedly installed on the inner bottom of the detection box, the bottom of the cold force temperature box is fixedly installed on the inner bottom of the detection box, four second temperature sensors are provided, and the four second temperature sensors are respectively fixedly installed inside the denaturation temperature box, the annealing temperature box, the extension temperature box and the cold force temperature box, and the heating tube is fixedly installed inside the denaturation temperature box, the annealing temperature box and the extension temperature box.

[0013] Furthermore, the heat supply component also includes a stirring motor, a stirring rod and a stirring blade, wherein the base of the stirring motor is fixedly mounted on the top of the denaturation temperature box, the annealing temperature box, the extension temperature box and the cooling temperature box, respectively, and the stirring rod and the stirring blade are respectively located inside the denaturation temperature box, the annealing temperature box, the extension temperature box and the cooling temperature box, the top of the stirring rod is fixedly mounted on the rotating shaft of the stirring motor, and the stirring blade is fixedly mounted on both sides of the stirring rod.

[0014] Furthermore, the heat supply component also includes a semiconductor refrigeration sheet, a cooling sheet, a heat sink, an interlayer box, a cooling fan and a grille opening, wherein the semiconductor refrigeration sheet is fixedly mounted on the wall of the cold temperature box, the cooling sheet is located inside the cold temperature box, the cooling sheet is fixedly mounted on the cooling end of the semiconductor refrigeration sheet, the heat sink is fixedly mounted on the heating end of the semiconductor refrigeration sheet, the heat sink is located outside the cold temperature box, the interlayer box is fixedly mounted on the outer wall of the cold temperature box, the cooling fan is fixedly mounted on the interlayer box, and the grille opening is opened on the wall of the detection box, and the grille opening is directly opposite to the end of the interlayer box away from the heat sink fan.

[0015] Furthermore, the heat supply assembly also includes a denaturation liquid inlet pipe, an annealing liquid inlet pipe, an extension liquid inlet pipe, a cold force liquid inlet pipe, a liquid supply pump, a first electric valve and a mixing barrel, wherein one end of the denaturation liquid inlet pipe is fixedly mounted on the denaturation temperature box, the other end of the denaturation liquid inlet pipe is fixedly mounted on the mixing barrel, one end of the annealing liquid inlet pipe is fixedly mounted on the annealing temperature box, the other end of the annealing liquid inlet pipe is fixedly mounted on the mixing barrel, one end of the extension liquid inlet pipe is fixedly mounted on the extension temperature box, and the other end of the extension liquid inlet pipe is fixedly mounted on the mixing barrel. One end of the cold force liquid inlet pipe is fixedly mounted on the mixing cylinder, one end of the cold force liquid inlet pipe is fixedly mounted on the cold force temperature box, and the other end of the cold force liquid inlet pipe is fixedly mounted on the mixing cylinder. Four liquid supply pumps are provided, and the four liquid supply pumps are respectively fixedly mounted on the denaturation liquid inlet pipe, the annealing liquid inlet pipe, the extension liquid inlet pipe and the end of the cold force liquid inlet pipe away from the mixing cylinder. Four first electric valves are provided, and the four first electric valves are respectively fixedly mounted on the denaturation liquid inlet pipe, the annealing liquid inlet pipe, the extension liquid inlet pipe and the end of the cold force liquid inlet pipe close to the mixing cylinder.

[0016] Furthermore, the heat supply component also includes a bracket, a transmission rod, an impeller, a spoiler blade, a third temperature sensor, a liquid supply pipe and a liquid supply channel, wherein the bracket is fixedly mounted on both sides inside the mixing barrel, the two ends of the transmission rod are rotatably mounted in the bracket, the impeller is fixedly mounted on the transmission rod, the spoiler blade is fixedly mounted on the transmission rod, the third temperature sensor is fixedly mounted on the mixing barrel, the bottom of the liquid supply pipe is fixedly mounted on the mixing barrel, the top of the liquid supply pipe is fixedly mounted on the sample plate, the liquid supply channel is opened in the sample plate, one end of the liquid supply channel is connected to the top of the liquid supply pipe, and the other end of the liquid supply channel is connected to the top interface of the spiral channel.

[0017] Furthermore, the heat supply component also includes a liquid outlet channel, a main liquid pipe, a switching tube, a fourth temperature sensor, a denaturation liquid outlet pipe, an annealing liquid outlet pipe, an extension liquid outlet pipe, a cooling liquid outlet pipe and a second electric valve, wherein the liquid outlet channel is opened in the sample plate, the liquid outlet channel is connected to the bottom interface of the spiral channel, the top of the main liquid pipe is fixedly mounted on the end of the sample plate away from the liquid supply pipe, the bottom of the main liquid pipe is fixedly mounted on one end of the switching tube, the fourth temperature sensor is fixedly mounted on one end of the switching tube close to the main liquid pipe, one end of the denaturation liquid outlet pipe is fixedly mounted on the switching tube, and the denaturation outlet The other end of the liquid pipe is fixedly mounted on the denaturation temperature box, one end of the annealing liquid outlet pipe is fixedly mounted on the adapter tube, the other end of the annealing liquid outlet pipe is fixedly mounted on the annealing temperature box, one end of the extension liquid outlet pipe is fixedly mounted on the adapter tube, the other end of the extension liquid outlet pipe is fixedly mounted on the extension temperature box, one end of the cold force liquid outlet pipe is fixedly mounted on the adapter tube, the other end of the cold force liquid outlet pipe is fixedly mounted on the cold force temperature box, four second electric valves are provided, and the four second electric valves are respectively fixedly mounted on the denaturation liquid outlet pipe, the annealing liquid outlet pipe, the extension liquid outlet pipe and the cold force liquid outlet pipe.

[0018] Furthermore, the airtight component includes an air ring vacuum pump, a two-way air duct, a third electric valve and an air supply channel, wherein the pump seat of the air ring vacuum pump is fixedly installed on the inner bottom of the detection box, the bottom of the two-way air duct is fixedly installed on the air ring vacuum pump, the top of the two-way air duct is fixedly installed on the sample plate, the third electric valve is fixedly installed on the bottom of the two-way air duct, the air supply channel is opened in the sample plate, one end of the air supply channel is connected to the top of the two-way air duct, and the other end of the air supply channel is connected to the airtight rubber ring.

[0019] The beneficial effects of the present invention are: The present invention heats the temperature in the sample well of the sample plate by high-speed flowing silicone oil, and then uses a denaturation temperature box, an annealing temperature box and an extension temperature box to adjust the temperature change required for the detection steps of denaturation, annealing and extension, and a cold temperature box to adjust the temperature balance of the silicone oil in the denaturation temperature box, and quickly transition the denaturation temperature to the annealing temperature, and quickly reduce the temperature in the sample well. The high-speed flowing silicone oil can quickly convert the corresponding temperatures of denaturation, annealing and extension, thereby improving the reaction efficiency, reducing non-specific products, improving the data quality and adapting to high-throughput detection.

[0020] The present invention starts the air ring vacuum pump first, which draws outside air and flows the air into the two-way air duct, the air supply channel and the airtight rubber ring. The airtight rubber ring bulges, and the bulged airtight rubber ring is tightly attached to the outer wall of the single hole of the experimental well plate in the sample hole to play a sealing role and avoid temperature loss inside the sample hole. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation of the present invention. In the accompanying drawings: Figure 1 This is a schematic diagram of the overall structure of a carbapenem resistance gene detection device proposed by the present invention; Figure 2 A cross-sectional view of a detection box of a carbapenem resistance gene detection device proposed by the present invention; Figure 3 A schematic diagram of the structure of a denaturation temperature box of a carbapenem resistance gene detection device proposed by the present invention; Figure 4 A carbapenem resistance gene detection device proposed by the present invention Figure 3 A magnified image of point A; Figure 5 A cross-sectional view of a mixing cylinder of a carbapenem resistance gene detection device proposed by the present invention; Figure 6 A schematic diagram of the structure of a cold temperature box of a carbapenem resistance gene detection device proposed by the present invention; Figure 7 A carbapenem resistance gene detection device proposed by the present invention Figure 6 A magnified view of point B; Figure 8 A schematic diagram of a spiral channel of a carbapenem resistance gene detection device proposed by the present invention; Fig. 9 A carbapenem resistance gene detection device proposed by the present invention Figure 8 Enlarged view of point C; Fig.10 A schematic diagram of the structure of a sample plate of a carbapenem resistance gene detection device proposed by the present invention; Fig.11 A schematic diagram of the structure of a stirring blade of a carbapenem resistance gene detection device proposed by the present invention; Fig.12 This is a schematic structural diagram of a heat dissipation fan of a carbapenem resistance gene detection device proposed by the present invention.

[0022] In the figure: 1, detection box; 101, first signal receiving port; 102, second signal receiving port; 2, flip cover; 3, interlayer plate; 4, sample plate; 401, sample hole; 5, computer controller; 6, displacement assembly; 601, displacement motor; 602, threaded rod; 603, guide rod; 604, threaded sleeve; 605, guide slide; 7, displacement sensor; 8, fluorescence detector; 9, heat supply assembly; 901, spiral channel; 9 02, denaturation temperature box; 903, annealing temperature box; 904, extension temperature box; 905, cooling temperature box; 906, second temperature sensor; 907, heating tube; 908, stirring motor; 909, stirring rod; 9010, stirring blade; 9011, semiconductor cooling sheet; 9012, cooling sheet; 9013, heat sink; 9014, interlayer box; 9015, cooling fan; 9016, grille; 9017, denaturation Liquid inlet pipe; 9018, annealing liquid inlet pipe; 9019, extension liquid inlet pipe; 9020, cold force liquid inlet pipe; 9021, liquid supply pump; 9022, first electric valve; 9023, mixing cylinder; 9024, bracket; 9025, transmission rod; 9026, impeller; 9027, spoiler blade; 9028, third temperature sensor; 9029, liquid supply pipe; 9030, liquid supply channel; 9031, liquid outlet channel; 9032, main liquid Tube; 9033, adapter tube; 9034, fourth temperature sensor; 9035, denaturation liquid outlet pipe; 9036, annealing liquid outlet pipe; 9037, extension liquid outlet pipe; 9038, cold force liquid outlet pipe; 9039, second electric valve; 10, airtight component; 1001, airtight rubber ring; 1002, air ring vacuum pump; 1003, two-way air duct; 1004, third electric valve; 1005, air supply channel; 11, first temperature sensor. DETAILED DESCRIPTION

[0023] The present invention will now be described in further detail with reference to the accompanying drawings. These drawings are simplified schematic diagrams, which only illustrate the basic structure of the present invention in a schematic manner, and therefore only show the components related to the present invention.

[0024] Please refer to Figures 1 to 12The present invention provides a carbapenem resistance gene detection device, comprising a detection box 1, a flap plate 2, an interlayer plate 3, a sample plate 4, a fluorescence detection unit, a thermal cycle unit and a computer controller 5, wherein the corner end of the flap plate 2 is rotatably mounted on the top of the detection box 1, the interlayer plate 3 is fixedly mounted on the inner wall of the detection box 1, the mounting column of the sample plate 4 is fixedly mounted on the interlayer plate 3, the fluorescence detection unit is slidably mounted inside the detection box 1, the fluorescence detection unit is used to monitor the change of the fluorescence signal during the reaction, and the fluorescence intensity is proportional to the amount of PCR product, so that the amount of the starting template can be quantitatively analyzed, the thermal cycle unit is fixedly mounted on the inner bottom of the detection box 1, the thermal cycle unit is used to control the change of temperature, so that DNA undergoes a cycle process of three steps of denaturation, annealing and extension, and the computer controller 5 is fixedly mounted on the inner bottom of the detection box 1; a sample hole 401 is opened on the top of the sample plate 4, and a plurality of sample holes 401 are opened, and the plurality of sample holes 401 are arranged in a matrix, and a 98-well plate for QPCR experiment is placed in the sample hole 401 of the sample plate 4.

[0025] The fluorescence detection unit includes a displacement component 6, a displacement sensor 7 and a fluorescence detector 8, the displacement component 6 is installed on the inner top of the detection box 1, the displacement sensor 7 is fixedly installed on the outer wall of the fluorescence detector 8, and the fluorescence detector 8 is threadedly installed on the displacement component 6; the thermal circulation unit includes a heat supply component 9, an airtight component 10 and a first temperature sensor 11, wherein the bottom of the heat supply component 9 is fixedly installed on the inner bottom of the detection box 1, the airtight component 10 is fixedly installed on the inner bottom of the detection box 1, the first temperature sensor 11 is fixedly installed on the bottom of the sample plate 4, and a plurality of first temperature sensors 11 are provided, the first temperature sensors 11 match the number of sample holes 401, each first temperature sensor 11 corresponds to a sample hole 401, and the first temperature sensor 11 is located directly below the bottom of the sample hole 401.

[0026] The heat supply component 9 includes a spiral channel 901, which is opened inside the sample plate 4. There are multiple spiral channels 901, and the number of spiral channels 901 matches that of the sample holes 401. Each spiral channel 901 is sleeved on a corresponding sample hole 401, and the spiral channel 901 is wound around the outside of the sample hole 401; the airtight component 10 includes an airtight rubber ring 1001, which is fixedly installed on the inner top of the sample hole 401. There are multiple airtight rubber rings 1001, which match the number of the sample holes 401. Each airtight rubber ring 1001 corresponds to a sample hole 401. When the airtight rubber ring 1001 is bulged, the bulged airtight rubber ring 1001 is tightly attached to the outer wall of the experimental well plate in the sample hole 401.

[0027] Specifically, the displacement assembly 6 includes a displacement motor 601, a threaded rod 602 and a guide rod 603, wherein the displacement motor 601 is fixedly mounted on the inner wall of the detection box 1, the two ends of the threaded rod 602 are rotatably mounted on the inner top of the detection box 1, the threaded rod 602 is fixedly mounted on the rotating shaft of the displacement motor 601, the guide rod 603 is fixedly mounted on the inner top of the detection box 1, and the guide rod 603 is located directly above the threaded rod 602; the displacement assembly 6 also includes a threaded sleeve 604 and a guide slide cylinder 605, the threaded sleeve 604 is threadedly sleeved on the threaded rod 602, the threaded sleeve 604 is fixedly mounted on the two ends of the fluorescence detector 8, the guide slide cylinder 605 is slidably mounted on the guide rod 603, and the guide slide cylinder 605 is fixedly mounted on the two ends of the fluorescence detector 8.

[0028] More specifically, the heat supply assembly 9 also includes a denaturation temperature box 902, an annealing temperature box 903, an extension temperature box 904, a cold force temperature box 905, a second temperature sensor 906 and a heating tube 907, wherein the bottom of the denaturation temperature box 902 is fixedly mounted on the inner bottom of the detection box 1, the bottom of the annealing temperature box 903 is fixedly mounted on the inner bottom of the detection box 1, the bottom of the extension temperature box 904 is fixedly mounted on the inner bottom of the detection box 1, and the bottom of the cold force temperature box 905 is fixedly mounted on the inner bottom of the detection box 1. At the bottom, four second temperature sensors 906 are arranged, and the four second temperature sensors 906 are respectively fixedly installed inside the denaturation temperature box 902, the annealing temperature box 903, the extension temperature box 904 and the cooling temperature box 905. The heating tube 907 is fixedly installed inside the denaturation temperature box 902, the annealing temperature box 903 and the extension temperature box 904. Silicone oil is stored in the denaturation temperature box 902, the annealing temperature box 903, the extension temperature box 904 and the cooling temperature box 905, and the silicone oil is low-viscosity silicone oil.

[0029] The heat supply component 9 also includes a stirring motor 908, a stirring rod 909 and a stirring blade 9010, wherein the base of the stirring motor 908 is fixedly installed on the top of the denaturation temperature box 902, the annealing temperature box 903, the extension temperature box 904 and the cooling temperature box 905, respectively, the stirring rod 909 and the stirring blade 9010 are respectively located inside the denaturation temperature box 902, the annealing temperature box 903, the extension temperature box 904 and the cooling temperature box 905, the top of the stirring rod 909 is fixedly installed on the rotating shaft of the stirring motor 908, and the stirring blade 9010 is fixedly installed on both sides of the stirring rod 909.

[0030] The heat supply component 9 also includes a semiconductor refrigeration sheet 9011, a cooling sheet 9012, a heat sink 9013, an interlayer box 9014, a cooling fan 9015 and a grille opening 9016, wherein the semiconductor refrigeration sheet 9011 is fixedly mounted on the wall of the cold temperature box 905, the cooling sheet 9012 is located inside the cold temperature box 905, the cooling sheet 9012 is fixedly mounted on the cooling end of the semiconductor refrigeration sheet 9011, the heat sink 9013 is fixedly mounted on the heating end of the semiconductor refrigeration sheet 9011, the heat sink 9013 is located outside the cold temperature box 905, the interlayer box 9014 is fixedly mounted on the outer wall of the cold temperature box 905, the cooling fan 9015 is fixedly mounted on the interlayer box 9014, and the grille opening 9016 is opened on the wall of the detection box 1, and the grille opening 9016 is directly opposite to the end of the interlayer box 9014 away from the heat sink 9015.

[0031] The heat supply assembly 9 also includes a denaturation liquid inlet pipe 9017, an annealing liquid inlet pipe 9018, an extension liquid inlet pipe 9019, a cold force liquid inlet pipe 9020, a liquid supply pump 9021, a first electric valve 9022 and a mixing barrel 9023, wherein one end of the denaturation liquid inlet pipe 9017 is fixedly mounted on the denaturation temperature box 902, the other end of the denaturation liquid inlet pipe 9017 is fixedly mounted on the mixing barrel 9023, one end of the annealing liquid inlet pipe 9018 is fixedly mounted on the annealing temperature box 903, the other end of the annealing liquid inlet pipe 9018 is fixedly mounted on the mixing barrel 9023, one end of the extension liquid inlet pipe 9019 is fixedly mounted on the extension temperature box 904, and the other end of the extension liquid inlet pipe 9019 is fixedly mounted on the mixing barrel 9023. On the mixing barrel 9023, one end of the cold force liquid inlet pipe 9020 is fixedly mounted on the cold force temperature box 905, and the other end of the cold force liquid inlet pipe 9020 is fixedly mounted on the mixing barrel 9023. Four liquid supply pumps 9021 are provided, and the four liquid supply pumps 9021 are respectively fixedly mounted on the denaturation liquid inlet pipe 9017, the annealing liquid inlet pipe 9018, the extended liquid inlet pipe 9019 and the cold force liquid inlet pipe 9020 at one end away from the mixing barrel 9023. Four first electric valves 9022 are provided, and the four first electric valves 9022 are respectively fixedly mounted on the denaturation liquid inlet pipe 9017, the annealing liquid inlet pipe 9018, the extended liquid inlet pipe 9019 and the cold force liquid inlet pipe 9020 at one end close to the mixing barrel 9023.

[0032] The heat supply component 9 also includes a bracket 9024, a transmission rod 9025, an impeller 9026, a spoiler blade 9027, a third temperature sensor 9028, a liquid supply pipe 9029 and a liquid supply channel 9030, wherein the bracket 9024 is fixedly mounted on both sides of the inside of the mixing barrel 9023, both ends of the transmission rod 9025 are rotatably mounted in the bracket 9024, the impeller 9026 is fixedly mounted on the transmission rod 9025, the spoiler blade 9027 is fixedly mounted on the transmission rod 9025, the third temperature sensor 9028 is fixedly mounted on the mixing barrel 9023, the bottom of the liquid supply pipe 9029 is fixedly mounted on the mixing barrel 9023, the top of the liquid supply pipe 9029 is fixedly mounted on the sample plate 4, and the liquid supply channel 9030 is opened in the sample plate 4, one end of the liquid supply channel 9030 is connected to the top of the liquid supply pipe 9029, and the other end of the liquid supply channel 9030 is connected to the top interface of the spiral channel 901.

[0033] The heat supply component 9 also includes a liquid outlet channel 9031, a main liquid pipe 9032, a switching tube 9033, a fourth temperature sensor 9034, a denaturation liquid outlet pipe 9035, an annealing liquid outlet pipe 9036, an extension liquid outlet pipe 9037, a cooling liquid outlet pipe 9038 and a second electric valve 9039, wherein the liquid outlet channel 9031 is opened in the sample plate 4, the liquid outlet channel 9031 is connected to the bottom interface of the spiral channel 901, the top of the main liquid pipe 9032 is fixedly installed on the end of the sample plate 4 away from the liquid supply pipe 9029, the bottom of the main liquid pipe 9032 is fixedly installed on one end of the switching tube 9033, the fourth temperature sensor 9034 is fixedly installed on one end of the switching tube 9033 close to the main liquid pipe 9032, and one end of the denaturation liquid outlet pipe 9035 is fixedly installed on the switching tube 9033. The other end of the denaturation liquid outlet pipe 9035 is fixedly mounted on the denaturation temperature box 902, one end of the annealing liquid outlet pipe 9036 is fixedly mounted on the adapter tube 9033, and the other end of the annealing liquid outlet pipe 9036 is fixedly mounted on the annealing temperature box 903, one end of the extension liquid outlet pipe 9037 is fixedly mounted on the adapter tube 9033, and the other end of the extension liquid outlet pipe 9037 is fixedly mounted on the extension temperature box 904, one end of the cold force liquid outlet pipe 9038 is fixedly mounted on the adapter tube 9033, and the other end of the cold force liquid outlet pipe 9038 is fixedly mounted on the cold force temperature box 905, and four second electric valves 9039 are provided, and the four second electric valves 9039 are respectively fixedly mounted on the denaturation liquid outlet pipe 9035, the annealing liquid outlet pipe 9036, the extension liquid outlet pipe 9037 and the cold force liquid outlet pipe 9038.

[0034] More specifically, the airtight component 10 includes an air ring vacuum pump 1002, a two-way air duct 1003, a third electric valve 1004 and an air supply channel 1005, wherein the pump seat of the air ring vacuum pump 1002 is fixedly installed on the inner bottom of the detection box 1, the bottom of the two-way air duct 1003 is fixedly installed on the air ring vacuum pump 1002, the top of the two-way air duct 1003 is fixedly installed on the sample plate 4, the third electric valve 1004 is fixedly installed on the bottom of the two-way air duct 1003, the air supply channel 1005 is opened in the sample plate 4, one end of the air supply channel 1005 is connected to the top of the two-way air duct 1003, and the other end of the air supply channel 1005 is connected to the airtight rubber ring 1001.

[0035] Furthermore, the experimental environment is cleaned to avoid cross contamination, and dedicated work areas are used to handle nucleic acid extraction, PCR reaction system construction and amplification detection respectively; individuals wear appropriate protective equipment, such as laboratory coats, gloves and masks.

[0036] Collect relevant clinical samples, such as blood, urine, sputum, etc.; then use a suitable DNA extraction kit to extract total DNA from the samples. During the extraction process, the aseptic operation principles must be strictly followed to avoid contamination by exogenous DNA.

[0037] According to the kit instructions, mix appropriate amounts of DNA template, primers, probe, PCR Master Mix and nuclease-free water to prepare a PCR reaction system; note that each sample and negative and positive controls must be prepared separately.

[0038] Add the prepared PCR reaction mixture into a 96-well experimental plate, and add a specified volume of reaction solution to each well; be careful to be gentle when adding samples to avoid creating bubbles.

[0039] The first signal receiving port 101 and the second signal receiving port 102 are connected via a data cable, and then connected to a host computer via a data cable, and the gene detection device of the present invention is controlled by the computer, the flip plate 2 is opened, and then the 96-well plate is placed in the sample hole 401 of the sample plate 4, and the flip plate 2 is closed.

[0040] The displacement motor 601 is started, and the rotation of the displacement motor 601 drives the threaded rod 602 to rotate. The rotation of the threaded rod 602 drives the threaded sleeve 604, the guide slide cylinder 605 and the fluorescence detector 8 to move along the guide rod 603, and the fluorescence detector 8 moves to the top of the sample plate 4.

[0041] The sample plate 4 is heated by a thermal cycle unit, so that the DNA in the 96-well experimental plate undergoes a cycle process of denaturation, annealing and extension.

[0042] Specific temperatures and functions of denaturation, annealing and extension: Process 1: Denaturation: Temperature: usually 94-96°C, sometimes up to 98°C; Function: At this high temperature, the hydrogen bonds of double-stranded DNA are broken, causing the DNA double strands to separate into single strands. This process is to expose the target DNA sequence so that it can bind to the primer; Time: Generally 15-30 seconds.

[0043] Process 2: Annealing: Temperature: usually 55-65°C, depending on the Tm value (melting temperature) of the primers; Function: At this temperature, the primer binds to the complementary sequence on the single-stranded DNA template. The Tm value refers to the temperature at which the primer binds halfway to the template. The annealing temperature is usually set to 5-10°C lower than the Tm value of the primer; Time: generally 15-60 seconds.

[0044] Process 3: Extension Temperature: usually 72°C; Function: At this temperature, DNA polymerase (such as Taq polymerase) is most active and can efficiently synthesize new DNA chains. The polymerase starts from the 3' end of the primer and extends along the template chain to synthesize new DNA molecules; Time: Generally 1 minute / kb (kilobase), that is, 1 minute is required for every 1000 bases.

[0045] Denaturation, annealing and extension cycle temperature transition time: Denaturation to annealing was 2 seconds; annealing to extension was 2 seconds; extension to denaturation of the next cycle was 2 seconds.

[0046] The three-step cycle of denaturation, annealing and extension: The computer controls the heating tube 907 to heat the silicone oil inside the denaturation temperature box 902, the annealing temperature box 903 and the extension temperature box 904; the semiconductor refrigeration sheet 9011 is started, and the cold force is transmitted to the cooling sheet 9012 through the semiconductor refrigeration sheet 9011, and the cooling sheet 9012 distributes the cold force to the silicone oil in the cooling temperature box 905, and then the stirring motor 908 is started, and the rotation of the rotating shaft of the stirring motor 908 drives the rotation of the stirring rod 909, and the rotation of the stirring rod 909 drives the rotation of the stirring blade 9010, and the rotation of the stirring blade 9010 disturbs the internal silicone oil of the denaturation temperature box 902, the annealing temperature box 903, the extension temperature box 904 and the cooling temperature box 905, so that the temperature inside them is more uniform.

[0047] The silicone oil temperature of the denaturation temperature box 902 is maintained at 98°C, the silicone oil temperature of the annealing temperature box 903 is maintained at 65°C, the silicone oil temperature of the extension temperature box 904 is maintained at 72°C, and the silicone oil temperature of the cold temperature box 905 is maintained at 0°C.

[0048] The gene detection device of the present invention first starts the air ring vacuum pump 1002, which draws external air and flows the air into the two-way air duct 1003, the air supply channel 1005 and the airtight rubber ring 1001. The airtight rubber ring 1001 swells, and the swollen airtight rubber ring 1001 is tightly attached to the outer wall of the single hole of the experimental well plate in the sample hole 401 to play a sealing role and avoid temperature loss inside the sample hole 401.

[0049] After the denaturation step, the liquid supply pump 9021 on the denaturation liquid inlet pipe 9017 is started, and the liquid supply pump 9021 extracts the silicone oil inside the denaturation temperature box 902 through the denaturation liquid inlet pipe 9017. The first electric valve 9022 of the denaturation liquid inlet pipe 9017 is opened (the first electric valve 9022 on the annealing liquid inlet pipe 9018, the extension liquid inlet pipe 9019 and the cooling liquid inlet pipe 9020 are closed), and the silicone oil in the denaturation liquid inlet pipe 9017 flows into the mixing barrel 9023. The oil enters the liquid supply pipe 9029, the liquid supply channel 9030, the spiral channel 901, the liquid outlet channel 9031, the main liquid pipe 9032, the adapter tube 9033 and the denaturation liquid outlet pipe 9035, the second electric valve 9039 of the denaturation liquid outlet pipe 9035 is opened (the second electric valves 9039 on the annealing liquid outlet pipe 9036, the extension liquid outlet pipe 9037 and the cooling liquid outlet pipe 9038 are closed), and the silicone oil flows back to the denaturation temperature box 902 through the denaturation liquid outlet pipe 9035. The temperature sensor 9028 detects the initial temperature of the silicone oil, and the fourth temperature sensor 9034 detects the temperature of the outflow of the silicone oil. When the temperature of the outflow of the silicone oil detected by the fourth temperature sensor 9034 reaches the range of 94-96°C, the liquid supply pump 9021 of the cold force temperature box 905 is turned on. The liquid supply pump 9021 allows the silicone oil in the cold force temperature box 905 to enter the mixing barrel 9023 through the annealing liquid inlet pipe 9018. The two silicone oils of different temperatures are mixed together in the mixing barrel 9023. The high-speed flow of the silicone oil drives the rotation of the impeller 9026. The rotation of the impeller 9026 drives the rotation of the turbulent blade 9027, so that the two silicone oils of different temperatures are fully mixed together. The third temperature sensor 9028 detects that the temperature of the mixed silicone oil is within the range of 94-96°C. The first temperature sensor 11 detects the temperature inside the sample hole 401 in real time, and controls the temperature inside the sample hole 401 by adjusting the flow rate of the silicone oil flowing out of the denaturation temperature box 902 and the cold force temperature box 905.

[0050] The temperature difference between denaturation and annealing is large. The annealing operation is to start the cold force temperature box 905 first, the liquid supply pump 9021 and the first electric valve 9022 of the cold force temperature box 905 are opened, and the silicone oil in the cold force temperature box 905 flows at a high speed to the cold force liquid inlet pipe 9020, the mixing cylinder 9023, the liquid supply pipe 9029, the liquid supply channel 9030, the spiral channel 901, the liquid outlet channel 9031, the main liquid pipe 9032, the adapter cylinder 9033 and the cold force liquid outlet pipe 9038, and then flows into the cold force temperature box 905 from the cold force liquid outlet pipe 9038. The first temperature sensor 11 detects After the temperature inside the sample hole 401 reaches 55-65°C, the silicone oil in the cooling temperature box 905 is disconnected, and then the annealing temperature box 903 is opened, and the liquid supply pump 9021 and the first electric valve 9022 of the annealing temperature box 903 are opened, and the silicone oil in the annealing temperature box 903 passes through the annealing liquid inlet pipe 9018, the mixing cylinder 9023, the liquid supply pipe 9029, the liquid supply channel 9030, the spiral channel 901, the liquid outlet channel 9031, the main liquid pipe 9032, the adapter cylinder 9033 and the annealing liquid outlet pipe 9036, and then flows back into the annealing temperature box 903 through the annealing liquid outlet pipe 9036.

[0051] The extension operation is to disconnect the silicone oil delivery of the extension liquid inlet pipe 9019, open the extension temperature box 904, open the liquid supply pump 9021 and the first electric valve 9022 of the extension temperature box 904, and the silicone oil in the extension temperature box 904 flows at a high speed to the extension liquid inlet pipe 9019, the mixing cylinder 9023, the liquid supply pipe 9029, the liquid supply channel 9030, the spiral channel 901, the liquid outlet channel 9031, the main liquid pipe 9032, the adapter cylinder 9033 and the extension liquid outlet pipe 9037, and then flows into the extension temperature box 904 from the extension liquid outlet pipe 9037.

[0052] The three steps of denaturation, annealing and extension were cycled 30-40 times.

[0053] The present invention heats the temperature in the sample hole 401 of the sample plate 4 by high-speed flowing silicone oil, and then uses the denaturation temperature box 902, annealing temperature box 903 and extension temperature box 904 to adjust the temperature changes required for the detection steps of denaturation, annealing and extension. The cold temperature box 905 regulates the temperature balance of the silicone oil in the denaturation temperature box 902, and the denaturation temperature quickly transitions to the annealing temperature, and quickly reduces the temperature in the sample hole 401. The high-speed flowing silicone oil can quickly convert the corresponding temperatures of denaturation, annealing and extension, thereby improving reaction efficiency, reducing non-specific products, improving data quality and adapting to high-throughput detection.

[0054] The benefits of shortening the transition temperature time between denaturation, annealing and extension: Benefit 1: Improve reaction efficiency: Shortened cycle time: Shortened temperature conversion time can significantly shorten the total time of each PCR cycle, thereby speeding up the entire PCR process; Increased throughput: In high-throughput assays, rapid temperature transitions can increase instrument throughput, allowing more reactions to be completed per unit time.

[0055] Benefit 2: Reduction of non-specific products: Rapid cooling: Rapidly cooling from the denaturation temperature to the annealing temperature can reduce the chance of nonspecific binding because the primers have less time to bind to non-complementary sequences; Rapid temperature ramp: Rapidly ramping from the annealing temperature to the extension temperature can reduce nonspecific amplification because the DNA polymerase has less time to synthesize DNA on nonspecifically bound primers.

[0056] Benefit 3: Improve data quality Consistency: Fast temperature conversion ensures more consistent temperature changes in each cycle, reducing errors caused by temperature fluctuations.

[0057] Precision: Rapid temperature transitions help maintain precise control of reaction conditions, improving the reproducibility and reliability of experimental results.

[0058] Benefit 4: Adaptable to high-throughput detection Automation: Rapid temperature conversion is suitable for automated high-throughput detection platforms, which can improve detection efficiency and accuracy; Save time: In clinical and research, fast temperature conversion can save a lot of time and improve work efficiency.

[0059] The computer controller 5 processes the data; analyzes the Ct value (threshold cycle number) through the qPCR software to determine whether the sample contains the target drug-resistant gene; the smaller the Ct value, the more initial copies of the target gene; checks the results of the negative control and the positive control to ensure the validity of the experiment; the negative control should not have an amplification signal, and the positive control should have an expected amplification curve; records the experimental results, including Ct values, amplification curves and other information; writes a test report based on the experimental results, which should include the experimental method, result interpretation and conclusion, etc., and is provided to the computer display screen for easy viewing by the staff.

[0060] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical scheme and inventive concept of the present invention within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

Claims

1. A carbapenem resistance gene detection device, characterized in that: The invention comprises a detection box (1), a flip plate (2), an interlayer plate (3), a sample plate (4), a fluorescence detection unit, a thermal cycle unit and a computer controller (5), wherein the corner end of the flip plate (2) is rotatably mounted on the top of the detection box (1), the interlayer plate (3) is fixedly mounted on the inner wall of the detection box (1), the mounting column of the sample plate (4) is fixedly mounted on the interlayer plate (3), the fluorescence detection unit is slidably mounted inside the detection box (1), the thermal cycle unit is fixedly mounted on the inner bottom of the detection box (1), and the computer controller (5) is fixedly mounted on the inner bottom of the detection box (1); A sample hole (401) is provided on the top of the sample plate (4), a plurality of the sample holes (401) are provided, and the plurality of the sample holes (401) are arranged in a matrix; The fluorescence detection unit comprises a displacement component (6), a displacement sensor (7) and a fluorescence detector (8); the displacement component (6) is mounted on the inner top of the detection box (1); the displacement sensor (7) is fixedly mounted on the outer wall of the fluorescence detector (8); and the fluorescence detector (8) is threadedly mounted on the displacement component (6); The thermal cycle unit comprises a heat supply component (9), an airtight component (10) and a first temperature sensor (11), wherein the bottom of the heat supply component (9) is fixedly mounted on the inner bottom of the detection box (1), the airtight component (10) is fixedly mounted on the inner bottom of the detection box (1), the first temperature sensor (11) is fixedly mounted on the bottom of the sample plate (4), and a plurality of first temperature sensors (11) are provided, the number of the first temperature sensors (11) matches the number of sample holes (401), each of the first temperature sensors (11) corresponds to a sample hole (401), and the first temperature sensor (11) is located directly below the bottom of the sample hole (401); The heat supply component (9) comprises a spiral channel (901), the spiral channel (901) being opened inside the sample plate (4), a plurality of the spiral channels (901) being opened, the number of the spiral channels (901) matching the number of the sample holes (401), and each of the spiral channels (901) being sleeved on a corresponding sample hole (401); The airtight component (10) comprises an airtight rubber ring (1001), wherein the airtight rubber ring (1001) is fixedly mounted on the inner top of the sample hole (401), and a plurality of the airtight rubber rings (1001) are provided, wherein the number of the airtight rubber rings (1001) matches the number of the sample holes (401), and each of the airtight rubber rings (1001) corresponds to a sample hole (401).

2. A carbapenem resistance gene detection device according to claim 1, characterized in that: The outer wall of the detection box (1) is provided with a first signal receiving port (101), and the outer wall of the detection box (1) is provided with a second signal receiving port (102).

3. A carbapenem resistance gene detection device according to claim 1, characterized in that: The displacement assembly (6) comprises a displacement motor (601), a threaded rod (602) and a guide rod (603), wherein the displacement motor (601) is fixedly mounted on the inner wall of the detection box (1), both ends of the threaded rod (602) are rotatably mounted on the inner top of the detection box (1), the threaded rod (602) is fixedly mounted on the rotating shaft of the displacement motor (601), and the guide rod (603) is fixedly mounted on the inner top of the detection box (1), and the guide rod (603) is located directly above the threaded rod (602); The displacement assembly (6) further comprises a threaded sleeve (604) and a guide slide cylinder (605), wherein the threaded sleeve (604) is threadedly sleeved on the threaded rod (602), wherein the threaded sleeve (604) is fixedly mounted on both ends of the fluorescence detector (8), and the guide slide cylinder (605) is slidably mounted on the guide rod (603), wherein the guide slide cylinder (605) is fixedly mounted on both ends of the fluorescence detector (8).

4. A carbapenem resistance gene detection device according to claim 1, characterized in that: The heat supply component (9) further comprises a denaturation temperature box (902), an annealing temperature box (903), an extension temperature box (904), a cold force temperature box (905), a second temperature sensor (906) and a heating tube (907), wherein the bottom of the denaturation temperature box (902) is fixedly mounted on the inner bottom of the detection box (1), the bottom of the annealing temperature box (903) is fixedly mounted on the inner bottom of the detection box (1), and the bottom of the extension temperature box (904) is fixedly mounted on the inner bottom of the detection box (1). The bottom of the cold force temperature box (905) is fixedly installed on the inner bottom of the detection box (1); four second temperature sensors (906) are provided, and the four second temperature sensors (906) are respectively fixedly installed inside the denaturation temperature box (902), the annealing temperature box (903), the extension temperature box (904) and the cold force temperature box (905); the heating tube (907) is fixedly installed inside the denaturation temperature box (902), the annealing temperature box (903) and the extension temperature box (904).

5. A carbapenem resistance gene detection device according to claim 4, characterized in that: The heat supply component (9) further comprises a stirring motor (908), a stirring rod (909) and a stirring blade (9010), wherein the base of the stirring motor (908) is fixedly mounted on the top of the denaturation temperature box (902), the annealing temperature box (903), the extension temperature box (904) and the cooling temperature box (905), respectively; the stirring rod (909) and the stirring blade (9010) are respectively located inside the denaturation temperature box (902), the annealing temperature box (903), the extension temperature box (904) and the cooling temperature box (905), the top of the stirring rod (909) is fixedly mounted on the rotating shaft of the stirring motor (908), and the stirring blade (9010) is fixedly mounted on both sides of the stirring rod (909).

6. A carbapenem resistance gene detection device according to claim 5, characterized in that: The heat supply component (9) further comprises a semiconductor refrigeration sheet (9011), a cooling sheet (9012), a heat sink (9013), an interlayer box (9014), a heat sink fan (9015) and a grille opening (9016), wherein the semiconductor refrigeration sheet (9011) is fixedly mounted on the wall of the cold temperature box (905), the cooling sheet (9012) is located inside the cold temperature box (905), the cooling sheet (9012) is fixedly mounted on the cooling end of the semiconductor refrigeration sheet (9011), and the heat sink (9013) is fixedly mounted on the heating end of the semiconductor refrigeration plate (9011), the heat sink (9013) is located on the outside of the cold temperature box (905), the interlayer box (9014) is fixedly mounted on the outer wall of the cold temperature box (905), the heat dissipation fan (9015) is fixedly mounted on the interlayer box (9014), the grille opening (9016) is opened on the box wall of the detection box (1), and the grille opening (9016) is directly opposite to the end of the interlayer box (9014) away from the heat dissipation fan (9015).

7. A carbapenem resistance gene detection device according to claim 6, characterized in that: The heat supply assembly (9) further comprises a denaturation liquid inlet pipe (9017), an annealing liquid inlet pipe (9018), an extension liquid inlet pipe (9019), a cooling liquid inlet pipe (9020), a liquid supply pump (9021), a first electric valve (9022) and a mixing barrel (9023), wherein one end of the denaturation liquid inlet pipe (9017) is fixedly mounted on the denaturation temperature box (902), the other end of the denaturation liquid inlet pipe (9017) is fixedly mounted on the mixing barrel (9023), one end of the annealing liquid inlet pipe (9018) is fixedly mounted on the annealing temperature box (903), the other end of the annealing liquid inlet pipe (9018) is fixedly mounted on the mixing barrel (9023), one end of the extension liquid inlet pipe (9019) is fixedly mounted on the extension temperature box (904), and the other end of the extension liquid inlet pipe (9019) is fixedly mounted On the mixing barrel (9023), one end of the cold force liquid inlet pipe (9020) is fixedly mounted on the cold force temperature box (905), and the other end of the cold force liquid inlet pipe (9020) is fixedly mounted on the mixing barrel (9023). Four liquid supply pumps (9021) are provided, and the four liquid supply pumps (9021) are respectively fixedly mounted on the end of the degeneration liquid inlet pipe (9017), the annealing liquid inlet pipe (9018), the extension liquid inlet pipe (9019), and the cold force liquid inlet pipe (9020) away from the mixing barrel (9023). Four first electric valves (9022) are provided, and the four first electric valves (9022) are respectively fixedly mounted on the end of the degeneration liquid inlet pipe (9017), the annealing liquid inlet pipe (9018), the extension liquid inlet pipe (9019), and the cold force liquid inlet pipe (9020) close to the mixing barrel (9023).

8. A carbapenem resistance gene detection device according to claim 7, characterized in that: The heat supply component (9) further comprises a bracket (9024), a transmission rod (9025), an impeller (9026), a spoiler blade (9027), a third temperature sensor (9028), a liquid supply pipe (9029) and a liquid supply channel (9030), wherein the bracket (9024) is fixedly mounted on both sides of the interior of the mixing barrel (9023), both ends of the transmission rod (9025) are rotatably mounted in the bracket (9024), the impeller (9026) is fixedly mounted on the transmission rod (9025), and the spoiler blade (9027) is fixedly mounted On the transmission rod (9025), the third temperature sensor (9028) is fixedly mounted on the mixing barrel (9023), the bottom of the liquid supply tube (9029) is fixedly mounted on the mixing barrel (9023), the top of the liquid supply tube (9029) is fixedly mounted on the sample plate (4), the liquid supply channel (9030) is opened in the sample plate (4), one end of the liquid supply channel (9030) is connected to the top of the liquid supply tube (9029), and the other end of the liquid supply channel (9030) is connected to the top interface of the spiral channel (901).

9. The carbapenem resistance gene detection device according to claim 8, characterized in that: The heat supply component (9) further comprises a liquid outlet channel (9031), a main liquid pipe (9032), a transfer tube (9033), a fourth temperature sensor (9034), a denaturation liquid outlet pipe (9035), an annealing liquid outlet pipe (9036), an extension liquid outlet pipe (9037), a cooling liquid outlet pipe (9038) and a second electric valve (9039), wherein the liquid outlet channel (9031) is opened in the sample plate (4), and the liquid outlet channel (9031) and the spiral channel ( The top of the main liquid pipe (9032) is fixedly mounted on an end of the sample plate (4) away from the liquid supply pipe (9029), the bottom of the main liquid pipe (9032) is fixedly mounted on an end of the adapter tube (9033), the fourth temperature sensor (9034) is fixedly mounted on an end of the adapter tube (9033) close to the main liquid pipe (9032), and one end of the denatured liquid outlet pipe (9035) is fixedly mounted on the adapter tube (9033). The other end of the denaturation liquid outlet pipe (9035) is fixedly mounted on the denaturation temperature box (902), one end of the annealing liquid outlet pipe (9036) is fixedly mounted on the adapter tube (9033), the other end of the annealing liquid outlet pipe (9036) is fixedly mounted on the annealing temperature box (903), one end of the extension liquid outlet pipe (9037) is fixedly mounted on the adapter tube (9033), the other end of the extension liquid outlet pipe (9037) is fixedly mounted on the extension temperature box ( 904), one end of the cold force liquid outlet pipe (9038) is fixedly mounted on the adapter tube (9033), the other end of the cold force liquid outlet pipe (9038) is fixedly mounted on the cold force temperature box (905), four second electric valves (9039) are provided, and the four second electric valves (9039) are respectively fixedly mounted on the denaturation liquid outlet pipe (9035), the annealing liquid outlet pipe (9036), the extension liquid outlet pipe (9037) and the cold force liquid outlet pipe (9038).

10. The carbapenem resistance gene detection device according to claim 1, characterized in that: The airtight component (10) comprises an air ring vacuum pump (1002), a two-way air duct (1003), a third electric valve (1004) and an air supply channel (1005), wherein the pump seat of the air ring vacuum pump (1002) is fixedly mounted on the inner bottom of the detection box (1), the bottom of the two-way air duct (1003) is fixedly mounted on the air ring vacuum pump (1002), the top of the two-way air duct (1003) is fixedly mounted on the sample plate (4), the third electric valve (1004) is fixedly mounted on the bottom of the two-way air duct (1003), the air supply channel (1005) is opened in the sample plate (4), one end of the air supply channel (1005) is connected to the top of the two-way air duct (1003), and the other end of the air supply channel (1005) is connected to the airtight rubber ring (1001).

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