Gram-positive bacterium pathogen PCR detection device and detection method

Abstract

The invention belongs to the technical field of medical instruments, and particularly relates to a gram-positive bacterium pathogen PCR detection device which comprises an analyzer main body, a loading groove is formed in the analyzer main body, a plurality of loading cylinders are assembled in the loading groove, one end of the analyzer main body is rotationally connected with a heat cover shell, and the other end of the analyzer main body is rotationally connected with the heat cover shell. A first groove is formed in the lower end of the interior of the heat cover shell, and a heat conduction plate and a conductive heating film are fixed in the first groove. When the device is in a closed state, the heat conduction sleeve is driven to be attached to the PRC pipe through the compressed elastic elements, so that the device can load the PRC pipes of different heights, diversified monitoring requirements are met, meanwhile, the pressure target value is set, the resilience force of the elastic elements is sensed through the pressure sensing elements, the compression amount of the multiple elastic elements is independently adjusted, and the pressure monitoring accuracy is improved. The pressure applied to the PRC pipes with different heights is consistent, and the phenomena of sealing performance, PCR pipe deformation or cracking and the like are avoided.

Description

Technical Field

[0001] The present invention belongs to the technical field of medical devices, and particularly relates to a Gram-positive bacterial pathogen PCR detection device and a detection method. Background Art

[0002] As a core detection method in the field of molecular biology, the fluorescence quantitative polymerase chain reaction (qPCR) technology plays an irreplaceable role in pathogen rapid identification, gene expression analysis, infectious disease prevention and control, etc. Its principle is to realize the quantitative analysis of target nucleic acid by monitoring the dynamic change of fluorescence signal in real time during the DNA amplification process through a fluorescence quantitative PCR analyzer. Gram-positive bacteria, as common clinical pathogens, the detection accuracy directly affects the diagnosis and treatment efficiency of infectious diseases. However, existing fluorescence quantitative PCR analyzers have significant limitations in practical applications, especially in the structural design of the detection device and the adaptability of the detection method.

[0003] Currently, the mainstream fluorescence quantitative PCR analyzers generally adopt a fixed thermal cover structure. To ensure the close contact between the thermal cover and the PCR tube to avoid evaporation contamination, the equipment has strict restrictions on the height and model of the PCR tube. Each detection process requires the use of PCR tubes of the same height or the same model. This single adaptability results in the need to frequently replace special PCR tubes and recalibrate the equipment parameters when the laboratory detects different samples, significantly increasing the detection cycle. Some improved PCR analysis devices adopt a double-slot design, attempting to achieve compatibility with multiple models of PCR tubes through an independent temperature control module. However, this solution requires an additional configuration of two sets of heating systems and thermal cover components. Under the condition of the same equipment volume, the single-batch detection capacity is reduced by about 40%, which not only reduces the detection throughput but also increases the manufacturing cost. In addition, the double-slot structure also brings problems such as complex optical path calibration and high maintenance cost, and it is difficult to meet the requirements of efficient detection in experiments or primary medical institutions.

[0004] In summary, there is an irreconcilable contradiction between the adaptability and detection efficiency of existing fluorescence quantitative PCR devices. The traditional single-slot design cannot meet the diverse detection needs, while the double-slot solution achieves limited compatibility at the cost of sacrificing the detection capacity, and neither has fundamentally solved the technical conflict between the equipment structure and the diversity of detection samples. This application proposes a Gram-positive bacterial pathogen PCR detection device and a detection method, which provide a new solution for the rapid and accurate detection of Gram-positive bacterial pathogens while maintaining the advantage of high capacity of a single slot. Summary of the Invention

[0005] The object of the present invention is to provide a PCR detection device for Gram-positive bacterial pathogens. When the device is in a closed state, a compressed elastic element drives a heat-conducting sleeve to fit with a PRC tube, enabling the device to load PRC tubes of different heights and meeting diverse monitoring requirements. At the same time, by setting a pressure target value and using a pressure sensing element to sense the resilience of the elastic element, the compression amount of multiple elastic elements can be independently adjusted to ensure that the pressure applied to PRC tubes of different heights is consistent, avoiding problems such as sealing issues, deformation or cracking of the PCR tubes.

[0006] The technical solution adopted by the present invention is as follows: A PCR detection device for Gram-positive bacterial pathogens includes an analyzer main body. A loading groove is provided inside the analyzer main body, and a plurality of loading cylinders are assembled inside the loading groove. One end of the analyzer main body is rotatably connected to a hot cover housing. A first groove is provided at the lower end inside the hot cover housing, and a heat-conducting plate and a conductive heating film are fixed inside the first groove, and the conductive heating film is located above the heat-conducting plate. It further includes: A plurality of pressing components. A plurality of the pressing components are all assembled inside the heat-conducting plate, and there is a one-to-one correspondence between the plurality of pressing components and the plurality of loading cylinders. The pressing component includes a heat-conducting sleeve and an elastic element. The heat-conducting sleeve is slidably connected inside the heat-conducting plate, and the elastic element is assembled inside the heat-conducting sleeve; A plurality of adjusting components. A plurality of the adjusting components are all assembled inside the hot cover housing, and there is a one-to-one correspondence between the plurality of adjusting components and the plurality of pressing components; Among them, the elastic element is always in a compressed state, and through the elastic element, a plurality of heat-conducting sleeves can be respectively in close contact with the tops of PCR tubes of different heights.

[0007] In a preferred embodiment, the adjusting component includes a guiding sleeve, a driving motor, a threaded rod, and a bottom plate. The guiding sleeve is fixed to the upper end inside the first groove, and the guiding sleeve is slidably connected to the heat-conducting sleeve. The driving motor is fixed inside the hot cover housing and is located above the guiding sleeve. The threaded rod is fixed to the output end of the driving motor, and the threaded rod is rotatably connected to the guiding sleeve. The bottom plate is threadedly connected to the outside of the threaded rod. The bottom plate is adapted to the elastic element, and the bottom plate is slidably connected to the guiding sleeve.

[0008] In a preferred embodiment, a pressure sensing element is fixed to the lower end of the bottom plate, and the pressure sensing element is in contact with the elastic element.

[0009] In a preferred embodiment, an anti-detachment plate is fixed to the lower end of the threaded rod and is located inside the elastic element.

[0010] In a preferred embodiment, a guiding boss is provided inside the guiding sleeve, a second guiding groove is formed on the outer side of the bottom plate, a first guiding groove is formed on the outer side of the heat conducting sleeve, and the bottom plate is adapted to the second guiding groove and the first guiding groove respectively.

[0011] In a preferred embodiment, an avoidance groove is formed on the outer side of the guiding sleeve, and the avoidance groove is adapted to the pressure sensing element.

[0012] In a preferred embodiment, a second groove is formed at the upper end inside the heat cover housing, a protective plate is fixed to the upper end of the second groove, and the driving motor is fixed inside the second groove.

[0013] In a preferred embodiment, the elastic element is a helical spring.

[0014] In a preferred embodiment, the elastic element is made of any one of the following materials: 302 stainless steel, chrome-silicon alloy steel, nickel-titanium alloy.

[0015] A detection method for a Gram-positive bacterial pathogen PCR detection device, applicable to the Gram-positive bacterial pathogen PCR detection device described in any one of the above, includes the following steps: St1: Add raw materials such as DNA template, primers, and enzymes to the corresponding PCR tubes, and then load the PCR tubes inside the loading cylinder. St2: Set the pressure target value, change the device from the open state to the closed state, and adjust the compression amount of each elastic element through the adjustment component. St3: Start the device to amplify the DNA.

[0016] The technical effects achieved by the present invention are: When the present invention loads PCR tubes with different heights inside multiple loading cylinders and the device is in the closed state, the elastic element in the compressed state drives the heat conducting sleeve to fit with the top of the PCR tube, and the PCR tube is pressed and sealed through the heat conducting sleeve, so that the device can load PCR tubes with different heights while maintaining the advantage of high capacity per slot, meeting diverse monitoring requirements, and improving the application range of the device.

[0017] By setting the pressure target value, and sensing the resilience of the elastic element through the pressure sensing element, starting the driving motor and driving the bottom plate and the pressure sensing element to move according to the resilience of the elastic element, the compression amount of each elastic element is independently adjusted, so that the device can independently adjust the compression amount of each elastic element for PCR tubes with different heights, ensuring that the pressure applied to each PCR tube is consistent, thus avoiding the problems of poor sealing of the PCR tube and evaporation of the reaction solution caused by insufficient pressure, and also preventing the deformation or cracking of the PCR tube caused by excessive pressure. Brief Description of the Drawings

[0018] Figure 1 is a schematic structural view of the whole of the present invention; Figure 2 is a schematic structural view of the opening and closing state of the whole structure of the present invention; Figure 3 is an exploded schematic structural view of the inside of the hot cover housing of the present invention; Figure 4 is a schematic structural view of the pressing component and the adjusting component of the present invention; Figure 5 is a cross-sectional view of the pressing component and the adjusting component of the present invention; Figure 6 is an exploded schematic structural view of the pressing component and the adjusting component of the present invention.

[0019] In the drawings, the list of components represented by each reference numeral is as follows: 10, analyzer main body; 11, loading groove; 12, loading cylinder; 13, hot cover housing; 14, first groove; 15, heat conducting plate; 16, conductive heating film; 17, second groove; 18, protection plate; 20, pressing component; 21, heat conducting sleeve; 22, elastic element; 23, first guiding groove; 30, adjusting component; 31, guiding sleeve; 32, driving motor; 33, threaded rod; 34, bottom plate; 35, pressure sensing element; 36, anti-disengagement plate; 37, guiding boss; 38, second guiding groove; 39, avoiding groove. Detailed Description of the Invention

[0020] In order to make the above objects, features and advantages of the present invention more obvious and understandable, the following detailed description of the specific embodiments of the present invention will be made in conjunction with the accompanying drawings of the specification.

[0021] In the following description, many specific details are set forth in order to fully understand the present invention. However, the present invention can also be implemented in other ways different from those described herein. Those skilled in the art can make similar generalizations without departing from the connotation of the present invention. Therefore, the present invention is not limited by the specific embodiments disclosed below.

[0022] Secondly, the so-called "one embodiment" or "embodiment" herein refers to a specific feature, structure or characteristic that can be included in at least one implementation of the present invention. The phrase "in a preferred embodiment" that appears in different places in this specification does not necessarily refer to the same embodiment, nor is it a separate or alternative embodiment that excludes other embodiments.

[0023] Next, the present invention will be described in detail with reference to the schematic diagrams. When describing the embodiments of the present invention in detail, for the convenience of explanation, the cross-sectional views showing the device structure will be enlarged locally not in accordance with the general scale, and the schematic diagrams are only examples and should not limit the scope of protection of the present invention herein. In addition, in actual production, three-dimensional spatial dimensions including length, width, and depth should be included.

[0024] Embodiment 1 Please refer to the attached Figures 1 to 3 As shown, this is the first embodiment of the present invention. This embodiment provides a PCR detection device for Gram-positive bacterial pathogens, including an analyzer main body 10. A loading groove 11 is provided inside the analyzer main body 10, and a plurality of loading cylinders 12 are assembled inside the loading groove 11. One end of the top of the analyzer main body 10 is rotatably connected to a hot cover housing 13. A first groove 14 is provided at the lower end inside the hot cover housing 13. A heat conducting plate 15 and a conductive heating film 16 are fixed inside the first groove 14, and the conductive heating film 16 is located above the heat conducting plate 15. It further includes: A plurality of pressing components 20, and a plurality of pressing components 20 are all assembled inside the heat conducting plate 15, and there is a one-to-one correspondence between the plurality of pressing components 20 and the plurality of loading cylinders 12. The pressing component 20 includes a heat conducting sleeve 21 and an elastic element 22. The heat conducting sleeve 21 is slidably connected inside the heat conducting plate 15, and the elastic element 22 is assembled inside the heat conducting sleeve 21; A plurality of adjusting components 30, and a plurality of adjusting components 30 are all assembled inside the hot cover housing 13, and there is a one-to-one correspondence between the plurality of adjusting components 30 and the plurality of pressing components 20; Among them, the elastic element 22 is always in a compressed state, and through the elastic element 22, the plurality of heat conducting sleeves 21 can be respectively in close contact with the tops of PCR tubes at different heights.

[0025] Furthermore, a locking element is assembled inside the analyzer main body 10, and a locking tongue element is assembled inside the hot cover housing 13. The locking element and the locking tongue element are adapted to each other. The locking element can limit the locking tongue element. When the locking element limits the locking tongue element, the device is in a closed state. When the locking element no longer limits the locking tongue element, the device is in an open state. When the device is in a closed state, through the cooperation of the analyzer main body 10 and the hot cover housing 13, the loading groove 11 is in a sealed state.

[0026] Specifically, when the device is in a closed state and no PRC tube is loaded inside it, the distance in the vertical direction between the bottom of the heat conducting sleeve 21 and the top of the loading cylinder 12 is denoted as L1. After the PRC tube is loaded inside the device, the distance in the vertical direction between the top of the PRC tube and the top of the loading cylinder 12 is denoted as L2, and L2 > L1.

[0027] It should be noted that inside the analyzer main body 10, there is at least an integrated control module, temperature control module, optoelectronic module, and power supply module. Among them, the control module is used to control the operation of each module of the system, realizing functions such as temperature rise and fall control, movement of mechanical components, and optoelectronic detection; at the same time, it monitors the status of each module and obtains detection data, mainly including the main board with a main control chip, temperature control board, mechanical control board, etc.; the temperature control module can provide a suitable temperature and temperature cycling environment for nucleic acid amplification and its processing and analysis, usually including a temperature rise and fall device, temperature sensor, radiator, and other auxiliary components, etc. The temperature control module can heat the PCR tubes inside the loading groove 11 and inside the loading cylinder 12. At the same time, the temperature control module and the conductive heating film 16 are electrically connected through wires. After the conductive heating film 16 is powered on, it can release heat. After the heat conduction plate 15 absorbs the heat released by the conductive heating film 16, it can heat up and transfer the heat to the heat conduction sleeve 21, causing the heat conduction sleeve 21 to heat up; the optoelectronic module can excite, conduct, convert, and collect fluorescence signals according to the program settings, generally consisting of an excitation light source, an optical path system, an optoelectronic converter, a signal amplifier, a detector, etc.; the power supply module can provide the power required for the operation of various modules inside the device, consisting of components such as an adapter and a switching power supply. Specifically, the fluorescence quantitative PCR analyzer is a mature existing application. The control module, temperature control module, optoelectronic module, power supply module, and other related components are not drawn in the embodiment. Their specific structures can refer to the prior art, and their installation positions can be adjusted adaptively according to actual needs. Here, no further elaboration will be made.

[0028] In this embodiment, when amplifying or detecting DNA, according to the detection requirements, raw materials such as DNA templates, primers, and enzymes are added to the corresponding PCR tubes. When the heights of the PCR tubes are different, the PCR tubes with different heights are respectively placed inside a plurality of loading cylinders 12. The hot cover housing 13 is rotated so that the hot cover housing 13 rotates towards the direction close to the loading cylinder 12, causing the device to change from an open state to a closed state. When the plurality of heat conduction sleeves 21 are respectively in contact with the PCR tubes with different heights, the PCR tubes push the heat conduction sleeves 21 to move upward, compressing the elastic elements 22 through the heat conduction sleeves 21. The PCR tubes with different heights can push the heat conduction sleeves 21 that fit them to different heights. Since the elastic elements 22 are in a compressed state, driven by the elastic elements 22, the heat conduction sleeves 21 can always be closely attached to the lids on the tops of the PCR tubes. The device is started, and the raw materials in the PCR tubes are circularly heated through the temperature control module, and the temperature control module causes the conductive heating film 16 to release heat. The heat conduction plate 15 is heated by the conductive heating film 16, and the heat absorption heat conduction plate 15 transfers the heat to the heat conduction sleeves 21, causing the heat conduction sleeves 21 to heat up, avoiding the condensation phenomenon of the reaction solution in the PCR tubes. Through the above scheme, each PCR tube independently corresponds to a heat conduction sleeve 21, and the lid of the PCR tube is pressed through the heat conduction sleeve 21 to maintain the airtightness of the PCR tube. On the premise of maintaining the advantage of a single slot with high capacity, the device can load PCR tubes with different heights, can meet diverse monitoring requirements, improves the applicable range of the device, and fundamentally solves the technical conflict between the equipment structure and the diversity of detection samples.

[0029] It should be noted that when amplifying (circularly heating) DNA, the process of a single cycle is denaturation, annealing, and extension. During the denaturation process, the temperature inside the loading groove 11 is 94 - 98 °C, and the temperature of the heat conduction sleeve 21 is 104 - 108 °C, avoiding the evaporation of the reaction solution (raw materials) in the PCR tubes and the condensation phenomenon occurring with the lids on the PCR tubes.

[0030] Secondly, please refer to again Figures 3 to 6As shown, the adjusting assembly 30 includes a guiding sleeve 31, a driving motor 32, a threaded rod 33, and a bottom plate 34. The guiding sleeve 31 is fixed to the upper end inside the first groove 14, and the guiding sleeve 31 is slidably connected to the heat conducting sleeve 21. The driving motor 32 is fixed inside the heat cover housing 13 and is located above the guiding sleeve 31, and the output end of the driving motor 32 extends into the guiding sleeve 31. The threaded rod 33 is fixed to the output end of the driving motor 32. The lower end of the outer side of the threaded rod 33 is located inside the elastic element 22, and the threaded rod 33 is rotatably connected to the guiding sleeve 31 through a sealed bearing. The bottom plate 34 is threadedly connected to the outer side of the threaded rod 33. The bottom plate 34 is adapted to the elastic element 22, and the bottom plate 34 is slidably connected to the guiding sleeve 31. After the driving motor 32 operates, the bottom plate 34 can adjust the compression amount of the elastic element 22.

[0031] Here, the guiding sleeve 31 and the control module are electrically connected by a wire. In this embodiment, the guiding sleeve 31 is preferably a servo motor.

[0032] In this embodiment, when amplifying DNA, PCR tubes of different heights are loaded inside the loading cylinder 12 according to the detection requirements. The heat cover housing 13 is rotated to change the device from the open state to the closed state. After the PCR tubes of different heights are in one-to-one contact with the multiple heat conducting sleeves 21, the PCR tubes push the heat conducting sleeves 21 to move upward, and the elastic element 22 is compressed through the heat conducting sleeves 21. Since the heights of the PCR tubes are different, the compression amounts of the elastic element 22 by the heat conducting sleeves 21 are also different (the compression amount of the elastic element 22 at the upper end of the PCR tube with a relatively low height is relatively small, and the compression amount of the elastic element 22 at the upper end of the PCR tube with a relatively high height is relatively large). When the compression amount of the elastic element 22 is relatively large, the elastic force applied by the elastic element 22 to the corresponding heat conducting sleeve 21 and the PCR tube is also relatively large, which is likely to cause the PCR tube to deform and crack. The corresponding driving motor 32 is started. Due to the fixed connection between the driving motor 32 and the threaded rod 33, the driving motor 32 drives the threaded rod 33 to rotate. Due to the fixed connection between the threaded rod 33 and the bottom plate 34 and the sliding connection between the bottom plate 34 and the guiding sleeve 31, the threaded rod 33 drives the bottom plate 34 to move upward. After the bottom plate 34 moves upward, the elastic element 22 with a relatively large compression amount can release an appropriate amount, thereby reducing the elastic force applied to the PCR tube and avoiding the PCR tube from deforming or cracking due to a large elastic force.

[0033] Next, please also refer to Figures 4 to 6 As shown, a pressure sensing element 35 is fixed to the lower end of the bottom plate 34, and the sensing end of the pressure sensing element 35 is in contact with the upper end of the elastic element 22.

[0034] Here, the pressure sensing element 35 and the control module are electrically connected by wires. At one end of the upper end of the analyzer main body 10 away from the thermal cover housing 13, a touch display component is provided, and the touch display component and the control module are electrically connected by wires. The pressure target value can be preset through the touch display component.

[0035] Furthermore, since the elastic element 22 is always in a compressed state, and the upper and lower ends of the elastic element 22 are respectively in contact with the sensing end of the pressure sensing element 35 and the heat conducting sleeve 21. During the detection process, the bottom of the heat conducting sleeve 21 is in close contact with the lid on the top of the PCR tube. The resilience force exerted by the compressed elastic element 22 on the heat conducting sleeve 21 is transmitted to the PCR tube through the heat conducting sleeve 21, that is, the pressing force exerted by the elastic element 22 on the PCR tube. During this process, the resilience force exerted by the compressed elastic element 22 on the sensing end of the pressure sensing element 35, the resilience force exerted on the heat conducting sleeve 21, and the pressing force exerted on the PCR tube are all equal.

[0036] In this embodiment, when amplifying DNA, the compression amounts of the elastic elements 22 corresponding to the PCR tubes with different heights are also different, and the pressing forces exerted by each elastic element 22 on the PCR tube are also different. The resilience force of the corresponding elastic element 22 is sensed by the pressure sensing element 35. When the compression amount of the elastic element 22 is too large and the resilience force is greater than the preset pressure target value, the driving motor 32 is started, and the driving motor 32 drives the bottom plate 34 and the pressure sensing element 35 to move upward, so that the elastic element 22 with too large a compression amount is released until the resilience force of the above elastic element 22 is equal to the pressure target value; on the contrary, when the compression amount of the elastic element 22 is too small and the resilience force is less than the preset pressure target value, the driving motor 32 is started, and the driving motor 32 drives the bottom plate 34 and the pressure sensing element 35 to move downward, so that the elastic element 22 with too small a compression amount is further compressed until the resilience force of the above elastic element 22 is equal to the pressure target value. Through the above scheme, the device can independently adjust the compression amount of each elastic element 22 for PCR tubes with different heights, ensuring that the pressure applied to each PCR tube is consistent. This not only avoids the problems of poor sealing of the PCR tube and evaporation of the reaction solution caused by insufficient pressure, but also prevents the deformation or cracking of the PCR tube caused by excessive pressure.

[0037] Secondly, please refer to again Figure 5 and Figure 6 , a retaining plate 36 is fixed at the lower end of the threaded rod 33 and inside the elastic element 22.

[0038] It should be noted that the pressure sensing element 35 is preferably an annular pressure sensor. A through hole is coaxially provided inside the annular pressure sensor. The diameter of the threaded rod 33 is denoted as D1, the inner diameter of the through hole is denoted as D2, and the outer diameter of the anti - detachment plate 36 is denoted as D3, where D3 > D2 > D1.

[0039] In this embodiment, when the driving motor 32 is started to drive the bottom plate 34 and the pressure sensing element 35 to move in the vertical direction, the setting of the anti - detachment plate 36 can prevent the detachment between the threaded rod 33 and the bottom plate 34, and between the threaded rod 33 and the pressure sensing element 35.

[0040] Please refer to again Figure 4 and Figure 6 , a guiding boss 37 is arranged inside the guiding sleeve 31, a second guiding groove 38 is formed on the outer side of the bottom plate 34, a first guiding groove 23 is formed on the outer side of the heat - conducting sleeve 21, and the bottom plate 34 is mutually adapted to the second guiding groove 38 and the first guiding groove 23. The guiding sleeve 31 and the bottom plate 34 are slidably connected through the cooperation of the guiding boss 37 and the second guiding groove 38, and the guiding sleeve 31 and the heat - conducting sleeve 21 are slidably connected through the cooperation of the guiding boss 37 and the first guiding groove 23.

[0041] In this embodiment, when adjusting the compression amount of the elastic element 22, the driving motor 32 is started to adjust the height of the bottom plate 34 and the pressure sensing element 35. Through the coordinated work of the guiding boss 37 and the second guiding groove 38, it is ensured that the guiding sleeve 31 can guide the bottom plate 34 during its movement and prevent the bottom plate 34 from rotating.

[0042] Please refer to again Figures 4 to 6 , an avoidance groove 39 is formed on the outer side of the guiding sleeve 31, and the avoidance groove 39 is adapted to the pressure sensing element 35.

[0043] In this embodiment, since the pressure sensing element 35 and the control module are electrically connected through a wire, when the pressure sensing element 35 moves, the connected wire will also move synchronously accordingly. The setting of the avoidance groove 39 is intended to ensure that the wire can move synchronously smoothly.

[0044] Please refer to again Figure 3 , a second groove 17 is formed at the upper end inside the heat - cover housing 13. A protective plate 18 is fixed at the upper end of the second groove 17. The driving motor 32 is fixed inside the second groove 17, and the second groove 17 is not communicated with the first groove 14.

[0045] In this embodiment, during the amplification of DNA, after the conductive heating film 16 releases heat, the internal temperature of the first groove 14 will increase synchronously. When denaturing DNA, the temperature inside the first groove 14 is as high as 104-108°C. By providing an independent second groove 17 for fixing the drive motor 32, it is possible to prevent the drive motor 32 from being in a high-temperature environment for a long time, thereby extending the service life of the drive motor 32 and enabling the device to work stably for a long time.

[0046] Here, a heat insulation layer (for example: ceramic fiber layer) is also assembled at the upper end inside the first groove 14.

[0047] It should be noted that a shaft hole is provided between the first groove 14 and the second groove 17, and the output end of the drive motor 32 extends into the guide sleeve 31 through the shaft hole. Since the output end of the drive motor 32 is inserted into the shaft hole and the top of the guide sleeve 31 can also prevent air convection between the first groove 14 and the second groove 17, therefore, in this application, the first groove 14 and the second groove 17 are described as not communicating.

[0048] In a preferred embodiment, the elastic element 22 is a helical spring, and the material of the elastic element 22 is any one of the following materials: 302 stainless steel, chrome-silicon alloy steel, nickel-titanium alloy, or other materials that can make springs and can be applicable to high-temperature environments above 120°C for a long time. In this embodiment, the material of the elastic element 22 is preferably 302 stainless steel.

[0049] In this embodiment, when denaturing DNA, the temperature inside the first groove 14 is as high as 104-108°C. Through the above settings, the elastic element 22 can work stably at high temperatures, avoiding the phenomenon of failure of the elastic element 22 caused by high temperatures.

[0050] Embodiment Two A detection method for a Gram-positive bacterial pathogen PCR detection device, applicable to a Gram-positive bacterial pathogen PCR detection device according to any one of Embodiment One, includes the following steps: St1: Add raw materials such as DNA templates, primers, and enzymes to the corresponding PCR tubes, and then load multiple PCR tubes into multiple loading cylinders 12 respectively; St2: Set the pressure target value, change the device from the open state to the closed state, sense the resilience of each elastic element 22 through the pressure sensing element 35, start the drive motor 32, and adjust the compression amount of each elastic element 22 according to the resilience of the elastic element 22, so that the pressing force applied by each elastic element 22 to the corresponding PRC tube is the same as the pressure target value; St3: Start the device to amplify or detect DNA.

[0051] Furthermore, when the device performs DNA amplification, it integrates real-time fluorescence quantitative PCR (qPCR) technology to achieve precise quantitative analysis of nucleic acids through dynamic monitoring of fluorescence signals. In a specific implementation, the optoelectronic module is built with a multi-channel fluorescence detection system, and the SYBR Green I fluorescence dye method or TaqMan probe method is used to track the amplification products in real time. After the device is started, the temperature control module drives the conductive heating film 16 and the heat conduction plate 15 to perform a temperature cycle according to a preset program (denaturation: 94-98°C, annealing: 55-65°C, extension: 72°C). At the same time, the optoelectronic module periodically excites the fluorescence signal in the PCR tube through an excitation light source (such as an LED). For the SYBR Green I method, the fluorescence intensity is proportional to the content of double-stranded DNA, and the signal is collected by the optoelectronic converter and then transmitted to the control module. If the TaqMan probe method is used, the probe is cleaved by Taq enzyme during amplification, the fluorophore is separated from the quencher, and the fluorescence signal increases with the accumulation of products. The software system built in the control module draws a fluorescence amplification curve in real time, and automatically calculates the initial template concentration by comparing the threshold cycle number (Ct value) with the standard curve. This process is visually displayed through the touch display component, and the user can directly obtain the quantitative result of the pathogen nucleic acid. In addition, the device supports multi-fluorescence channel detection (such as FAM, HEX, VIC), can analyze multiple target genes simultaneously, and meets the complex detection requirements such as genotyping of Gram-positive bacteria drug-resistant genes.

[0052] In a specific embodiment, this embodiment provides a method for detecting Gram-positive bacterial pathogens based on real-time fluorescence quantitative PCR (qPCR) technology. The specific steps are as follows: Step 1: Sample preparation and reaction system configuration Extract DNA or RNA from the sample to be tested, and use a reverse transcription kit (for RNA samples) to synthesize cDNA. Configure the PCR reaction solution in proportion, including the following components: 10×PCR buffer (containing Mg 2+ ) dNTP mixture (200 μm each) Forward / reverse primers (0.2 μm each) SYBR Green I fluorescence dye or TaqMan probe (0.1 μm) Taq DNA polymerase (1 U / μL) Template DNA / cDNA (2-5 μL) Step 2: Loading and parameter setting Dispense the configured reaction solution into PCR tubes and load them into the loading cylinder 12 of the analyzer main body 10. Set the qPCR program through the touch display component: Pre-denaturation: 95°C for 3 min Cyclic amplification (40 cycles): Denaturation at 95°C for 15 s → Annealing at 60°C for 30 s → Extension at 72°C for 30 s; Melting curve analysis: 95°C for 15 s → 60°C for 1 min → 95°C for 15 s (fluorescence signal continuously monitored); Step 3: Real-time fluorescence detection and data analysis After starting the device, the optoelectronic module collects the fluorescence signals of each cycle in real time, and the control module generates fluorescence amplification curves and melting curves. The Ct value is automatically calculated by software and compared with the standard curve to quantify the copy number of the target gene in the sample. If melting curve analysis is used, the specificity of the amplification product can also be verified by the characteristic melting temperature (Tm value). In this embodiment, by integrating the qPCR technology, highly sensitive and highly specific detection of Gram-positive bacterial pathogens is achieved. The multi-channel fluorescence synchronous acquisition function supports multiplex PCR detection, and multiple drug resistance genes (such as mecA, vanA) can be analyzed simultaneously, significantly improving the detection efficiency. Combining with the adaptive pressing component in Embodiment 1 ensures the sealing of PCR tubes of different specifications and avoids deviations in fluorescence signals caused by evaporation or condensation.

[0053] The working principle of the present invention is as follows: When setting the pressure target value through the touch display component and amplifying or detecting DNA, according to the detection requirements, raw materials such as DNA templates, primers, and enzymes are added to the corresponding PCR tubes, and then PCR tubes of different heights are respectively loaded inside a plurality of loading cylinders 12. The hot cover housing 13 is rotated to change the device from an open state to a closed state. When the plurality of heat conduction sleeves 21 are respectively in contact with PCR tubes of different heights, the PCR tubes push the heat conduction sleeves 21 to move upward, and the elastic elements 22 are compressed through the heat conduction sleeves 21. PCR tubes of different heights can push the heat conduction sleeves 21 that fit them to different heights. Driven by the elastic elements 22 in the compressed state, the heat conduction sleeves 21 can always be in close contact with the lids on the tops of the PCR tubes. At the same time, the compression amounts of the respective elastic elements 22 are also different. The pressure sensing element 35 senses the resilience of the corresponding elastic element 22. If the resilience of the elastic element 22 is not the same as the pressure target value, the drive motor 32 is started, and the drive motor 32 drives the bottom plate 34 and the pressure sensing element 35 to move in the vertical direction, thereby automatically adjusting the compression amount of the elastic element 22 until the resilience of the elastic element 22 is the same as the pressure target value. The device is started to amplify or detect DNA. Thus, on the premise of maintaining the advantage of high capacity per slot, the device can load PCR tubes of different heights, meet diverse monitoring requirements, improve the applicable range of the device, and can also independently adjust the compression amounts of the respective elastic elements 22 for PCR tubes of different heights to ensure that the pressure applied to each PCR tube is consistent. This not only avoids problems such as poor sealing of PCR tubes and evaporation of reaction solutions caused by insufficient pressure, but also prevents deformation or cracking of PCR tubes caused by excessive pressure.

[0054] The above is only the preferred embodiment of the present invention. It should be noted that for those of ordinary skill in the art in this technical field, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention. The structures, devices, and operation methods not specifically described and explained in the present invention, unless otherwise specified and limited, are implemented according to the conventional means in the art.

Claims

1. A PCR detection device for Gram-positive pathogens, characterized in that: The analyzer comprises an analyzer body (10), wherein a loading groove (11) is provided inside the analyzer body (10), and a plurality of loading cylinders (12) are mounted inside the loading groove (11), one end of the analyzer body (10) is rotatably connected to a heat cover shell (13), a first groove (14) is provided at the lower end inside the heat cover shell (13), a heat conducting plate (15) and a conductive heating film (16) are fixed inside the first groove (14), and the conductive heating film (16) is located at the upper end of the heat conducting plate (15), and further comprises: A plurality of press-fit assemblies (20), each of the plurality of press-fit assemblies (20) being assembled inside the heat-conducting plate (15), and each of the plurality of press-fit assemblies (20) and the plurality of loading cylinders (12) corresponding one to another, the press-fit assemblies (20) comprising a heat-conducting sleeve (21) and an elastic element (22), the heat-conducting sleeve (21) being slidably connected inside the heat-conducting plate (15), and the elastic element (22) being assembled inside the heat-conducting sleeve (21); A plurality of adjustment components (30), wherein the plurality of adjustment components (30) are all assembled inside the heat cover shell (13), and there is a one-to-one correspondence between the plurality of adjustment components (30) and the plurality of pressing components (20); The elastic element (22) is always in a compressed state, and the elastic element (22) can enable a plurality of heat-conducting sleeves (21) to fit tightly against the tops of PCR tubes at different heights.

2. A Gram-positive bacterial pathogen PCR detection device according to claim 1, characterized in that: The adjustment assembly (30) comprises a guide sleeve (31), a drive motor (32), a threaded rod (33) and a bottom plate (34); the guide sleeve (31) is fixed to the upper end inside the first groove (14), and the guide sleeve (31) and the heat-conducting sleeve (21) are slidably connected; the drive motor (32) is fixed inside the heat cover shell (13) and is located at the upper end of the guide sleeve (31); the threaded rod (33) is fixed to the output end of the drive motor (32), and the threaded rod (33) and the guide sleeve (31) are rotatably connected; the bottom plate (34) is threadedly connected to the outer side of the threaded rod (33); the bottom plate (34) and the elastic element (22) are matched, and the bottom plate (34) and the guide sleeve (31) are slidably connected.

3. A Gram-positive bacterial pathogen PCR detection device according to claim 2, characterized in that: A pressure sensing element (35) is fixed to the lower end of the bottom plate (34), and the pressure sensing element (35) and the elastic element (22) are in close contact with each other.

4. A Gram-positive bacterial pathogen PCR detection device according to claim 2, characterized in that: An anti-slip plate (36) is fixed to the lower end of the threaded rod (33).

5. A Gram-positive bacterial pathogen PCR detection device according to claim 2, characterized in that: A guide boss (37) is provided inside the guide sleeve (31), a second guide groove (38) is provided on the outside of the bottom plate (34), a first guide groove (23) is provided on the outside of the heat-conducting sleeve (21), and the bottom plate (34) and the second guide groove (38) as well as the bottom plate (34) and the first guide groove (23) are mutually adapted.

6. A Gram-positive bacterial pathogen PCR detection device according to claim 3, characterized in that: An escape groove (39) is provided on the outer side of the guide sleeve (31), and the escape groove (39) is adapted to the pressure sensing element (35).

7. A Gram-positive bacterial pathogen PCR detection device according to claim 2, characterized in that: A second groove (17) is provided at the upper end of the interior of the heat cover shell (13), a protective plate (18) is fixed to the upper end of the second groove (17), and the drive motor (32) is fixed inside the second groove (17).

8. A Gram-positive bacterial pathogen PCR detection device according to claim 1, characterized in that: The elastic element (22) is a coil spring.

9. A Gram-positive bacterial pathogen PCR detection device according to claim 8, characterized in that: The elastic element (22) is made of any one of the following materials: 302 stainless steel, chrome silicon alloy steel, and nickel titanium alloy.

10. A detection method of a Gram-positive bacterial pathogen PCR detection device, applicable to a Gram-positive bacterial pathogen PCR detection device according to any one of claims 1 to 9, characterized in that: The following steps are involved: St1: Add the raw materials into the corresponding PCR tube, and then load the PCR tube into the loading cylinder (12); St2: setting a target pressure value, changing the device from an open state to a closed state, and adjusting the compression amount of each elastic element (22) through an adjusting component (30); St3: Start the device to amplify DNA.

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