A PCR detection device and method for Gram-positive bacterial pathogens

Abstract

This invention belongs to the field of medical device technology, specifically relating to a PCR detection device for Gram-positive bacterial pathogens. The device includes an analyzer body with a loading groove inside. Multiple loading cylinders are assembled inside the loading groove. A heat-conducting cover is rotatably connected to one end of the analyzer body. A first groove is formed at the lower end of the heat-conducting cover, and a heat-conducting plate and a conductive heating film are fixed inside the first groove. When the device is in a closed state, the compressed elastic element drives the heat-conducting sleeve to fit against the PRC tube, allowing the device to load PRC tubes of different heights to meet diverse monitoring needs. Simultaneously, by setting a target pressure value, a pressure sensor detects the rebound force of the elastic element, independently adjusting the compression of multiple elastic elements to ensure consistent pressure applied to PRC tubes of different heights, avoiding sealing problems, PCR tube deformation, or cracking.

Description

Technical Field

[0001] This invention belongs to the field of medical device technology, specifically relating to a PCR detection device and method for Gram-positive bacterial pathogens. Background Technology

[0002] Quantitative real-time polymerase chain reaction (qPCR) technology, as a core detection method in molecular biology, plays an irreplaceable role in rapid pathogen identification, gene expression analysis, and infectious disease control. Its principle is to achieve quantitative analysis of target nucleic acids by monitoring the dynamic changes of fluorescence signals during DNA amplification in real time using a qPCR analyzer. Gram-positive bacteria are common clinical pathogens, and the accuracy of their detection directly affects the diagnostic and treatment efficiency of infectious diseases. However, existing qPCR analyzers have significant limitations in practical applications, particularly in the structural design of the detection device and its compatibility with the detection method.

[0003] Most mainstream real-time PCR analyzers currently employ a fixed heated cap structure. To ensure tight contact between the heated cap and the PCR tubes and prevent evaporation contamination, the equipment imposes strict limitations on the height and type of PCR tubes. Each testing session requires the use of PCR tubes of the same height or type. This single compatibility necessitates frequent replacement of dedicated PCR tubes and recalibration of equipment parameters when testing different samples, significantly increasing the testing cycle. Some improved PCR analyzers adopt a dual-chamber design, attempting to achieve compatibility with multiple PCR tube types through independent temperature control modules. However, this approach requires two additional heating systems and heated cap assemblies. With the same equipment volume, the single-batch testing capacity is reduced by approximately 40%, decreasing throughput and increasing manufacturing costs. Furthermore, the dual-chamber structure leads to complex optical path calibration and high maintenance costs, making it difficult to meet the high-efficiency testing needs of laboratories or primary healthcare institutions.

[0004] In summary, existing quantitative PCR devices suffer from an irreconcilable conflict between adaptability and detection efficiency. Traditional single-chamber designs cannot meet diverse detection needs, while dual-chamber solutions achieve limited compatibility at the expense of detection capacity; neither fundamentally resolves the technical conflict between device structure and the diversity of samples to be detected. This application proposes a PCR detection device and method for Gram-positive bacterial pathogens, providing a novel solution for rapid and accurate detection of Gram-positive bacterial pathogens while maintaining the high capacity advantage of single-chamber designs. Summary of the Invention

[0005] The purpose of this 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 against the PRC tube, allowing the device to accommodate PRC tubes of different heights to meet diverse monitoring needs. At the same time, by setting a target pressure value, a pressure sensing element senses the rebound force of the elastic element and independently adjusts the compression of multiple elastic elements to ensure consistent pressure applied to PRC tubes of different heights, avoiding sealing problems, PCR tube deformation, or cracking.

[0006] The specific technical solution adopted by this invention is as follows:

[0007] A PCR detection device for Gram-positive bacterial pathogens includes an analyzer body with a loading groove inside. Multiple loading cylinders are assembled inside the loading groove. A heat-sealed housing is rotatably connected to one end of the analyzer body. A first groove is formed at the lower end of the heat-sealed housing. A heat-conducting plate and a conductive heating film are fixed inside the first groove, with the conductive heating film located at the upper end of the heat-conducting plate. The device also includes:

[0008] Multiple pressing components are assembled inside the heat-conducting plate, and there is a one-to-one correspondence between the multiple pressing components and multiple loading cylinders. Each pressing component includes a heat-conducting sleeve and an elastic element. The heat-conducting sleeve is slidably connected to the inside of the heat-conducting plate, and the elastic element is assembled inside the heat-conducting sleeve.

[0009] Multiple adjustment components are assembled inside the heat cover housing, and there is a one-to-one correspondence between the multiple adjustment components and the multiple pressing components;

[0010] The elastic element is always in a compressed state, which allows multiple heat-conducting sleeves to fit tightly against the tops of PCR tubes at different heights.

[0011] In a preferred embodiment, the adjusting assembly includes a guide sleeve, a drive motor, a threaded rod, and a base plate. The guide sleeve is fixed to the upper end inside the first groove, and the guide sleeve and the heat-conducting sleeve are slidably connected. The drive motor is fixed inside the heat cover housing and located at the upper end of the guide sleeve. The threaded rod is fixed to the output end of the drive motor, and the threaded rod and the guide sleeve are rotatably connected. The base plate is threaded to the outside of the threaded rod. The base plate is adapted to the elastic element, and the base plate and the guide sleeve are slidably connected.

[0012] In a preferred embodiment, a pressure sensing element is fixed to the lower end of the base plate, and the pressure sensing element and the elastic element are in contact.

[0013] In a preferred embodiment, an anti-detachment plate is fixed at the lower end of the threaded rod, located inside the elastic element.

[0014] In a preferred embodiment, the guide sleeve has a guide boss inside, the base plate has a second guide groove on the outer side, the heat-conducting sleeve has a first guide groove on the outer side, and the base plate and the second guide groove, as well as the base plate and the first guide groove, are mutually compatible.

[0015] In a preferred embodiment, the outer side of the guide sleeve is provided with a clearance groove, and the clearance groove is adapted to the pressure sensing element.

[0016] In a preferred embodiment, a second groove is provided at the upper end of the interior of the heat cover housing, a protective plate is fixed at the upper end of the second groove, and the drive motor is fixed inside the second groove.

[0017] In a preferred embodiment, the elastic element is a helical spring.

[0018] In a preferred embodiment, the elastic element is made of any one of the following materials: 302 stainless steel, chromium-silicon alloy steel, or nickel-titanium alloy.

[0019] A detection method for a Gram-positive bacterial pathogen PCR detection device, applicable to any of the above-described Gram-positive bacterial pathogen PCR detection devices, comprising the following steps:

[0020] St1: Add the DNA template, primers, enzymes and other raw materials to the corresponding PCR tube, and then load the PCR tube into the loading tube;

[0021] St2: Set the target pressure value, change the device from the open / closed state to the closed state, and adjust the compression of each elastic element by adjusting the adjustment component;

[0022] St3: Start the device to amplify the DNA.

[0023] The technical effects achieved by this invention are as follows:

[0024] This invention loads PRC tubes of different heights inside multiple loading cylinders and the device is in a closed state. The compressed elastic element drives the heat-conducting sleeve to fit against the top of the PRC tube. The heat-conducting sleeve presses and seals the PRC tube, allowing the device to load PCR tubes of different heights while maintaining the high capacity advantage of a single tank. This meets diverse monitoring needs and improves the applicability of the device.

[0025] This invention sets a target pressure value and senses the rebound force of the elastic elements through a pressure sensing element. Based on the rebound force of the elastic elements, a drive motor is activated to move the base plate and the pressure sensing element, allowing independent adjustment of the compression of multiple elastic elements. This enables the device to independently adjust the compression of each elastic element for PCR tubes of different heights, ensuring consistent pressure applied to each PCR tube. This avoids problems such as poor PCR tube sealing and evaporation of reaction solution due to insufficient pressure, and also prevents PCR tube deformation or cracking due to excessive pressure. Attached Figure Description

[0026] Figure 1 This is a schematic diagram of the overall structure of the present invention;

[0027] Figure 2 This is a schematic diagram of the overall structure of the present invention in its open and closed state;

[0028] Figure 3 This is an exploded view of the internal structure of the heat-cap shell of the present invention;

[0029] Figure 4 This is a schematic diagram of the structure of the pressing component and the adjusting component of the present invention;

[0030] Figure 5 This is a structural cross-sectional view of the pressing assembly and adjusting assembly of the present invention;

[0031] Figure 6 This is an exploded view of the structure of the pressing component and the adjusting component of the present invention.

[0032] In the accompanying drawings, the components represented by the reference numerals are as follows:

[0033] 10. Analyzer body; 11. Loading groove; 12. Loading cylinder; 13. Heat cover shell; 14. First groove; 15. Heat-conducting plate; 16. Conductive heating film; 17. Second groove; 18. Protective plate;

[0034] 20. Pressing assembly;

[0035] 21. Thermally conductive sleeve; 22. Elastic element; 23. First guide groove;

[0036] 30. Adjustment components;

[0037] 31. Guide sleeve; 32. Drive motor; 33. Threaded rod; 34. Base plate; 35. Pressure sensing element; 36. Anti-detachment plate; 37. Guide boss; 38. Second guide groove; 39. Clearance groove. Detailed Implementation

[0038] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings.

[0039] Many specific details are set forth in the following description in order to provide a full understanding of the invention. However, the invention may also be practiced in other ways different from those described herein, and those skilled in the art can make similar extensions without departing from the spirit of the invention. Therefore, the invention is not limited to the specific embodiments disclosed below.

[0040] Secondly, the term "an embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The phrase "in a preferred embodiment" appearing in different places throughout this specification does not necessarily refer to the same embodiment, nor is it a single or selective embodiment that mutually excludes other embodiments.

[0041] Secondly, the present invention is described in detail with reference to the schematic diagrams. When detailing the embodiments of the present invention, for ease of explanation, the cross-sectional views illustrating the device structure may be partially enlarged, not according to the usual scale. Furthermore, the schematic diagrams are merely examples and should not limit the scope of protection of the present invention. In addition, actual fabrication should include three-dimensional spatial dimensions of length, width, and depth.

[0042] Example 1

[0043] Please see the appendix 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 body 10. The analyzer body 10 has a loading groove 11 inside, and multiple loading cylinders 12 are assembled inside the loading groove 11. A heat-conducting shell 13 is rotatably connected to one end of the top of the analyzer body 10. A first groove 14 is formed at the lower end of the heat-conducting 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 above the heat-conducting plate 15. The device also includes:

[0044] Multiple pressing components 20 are assembled inside the heat-conducting plate 15, and there is a one-to-one correspondence between the multiple pressing components 20 and the multiple 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 to the inside of the heat-conducting plate 15, and the elastic element 22 is assembled inside the heat-conducting sleeve 21.

[0045] Multiple adjustment components 30 are assembled inside the heat cover housing 13, and there is a one-to-one correspondence between the multiple adjustment components 30 and the multiple pressing components 20;

[0046] The elastic element 22 is always in a compressed state, and the elastic element 22 enables multiple heat-conducting sleeves 21 to fit tightly against the top of PCR tubes of different heights.

[0047] Furthermore, the analyzer body 10 is equipped with a locking element inside, and the hot cover housing 13 is equipped with a locking tongue element inside. The locking element and the locking tongue element are compatible with 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 or closed state. When the device is in a closed state, the loading groove 11 is sealed by the cooperation of the analyzer body 10 and the hot cover housing 13.

[0048] Specifically, when the device is in a closed state and no PRC tube is loaded inside it, the vertical distance 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 vertical distance between the top of the PRC tube and the top of the loading cylinder 12 is denoted as L2, where L2 > L1.

[0049] It should be noted that the analyzer body 10 also integrates at least a control module, a temperature control module, a photoelectric module, and a power supply module. The control module controls the operation of each module, implementing functions such as temperature control, movement of mechanical parts, and photoelectric detection; it also monitors the status of each module and acquires detection data. This module mainly includes a mainboard with a main control chip, a temperature control board, and a mechanical control board. The temperature control module provides a suitable temperature and temperature cycling environment for nucleic acid amplification and processing analysis. It typically includes a temperature control device, a temperature sensor, a heat sink, and other auxiliary components. The temperature control module heats the PCR tubes inside the loading groove 11 and inside the loading cylinder 12. Simultaneously, the temperature control module and the conductive heating film 16 are electrically connected via wires. After the conductive heating film 16 is energized... The heat-conducting plate 15 can release heat, and after absorbing the heat released by the conductive heating film 16, it can heat up and transfer the heat to the heat-conducting sleeve 21, causing the heat-conducting sleeve 21 to heat up. The photoelectric module can excite, conduct, convert and collect fluorescence signals according to the program settings. It is generally composed of an excitation light source, an optical path system, a photoelectric converter, a signal amplifier, a detector, etc. The power module can provide the power required for the operation of various modules inside the device. It is composed of components such as an adapter and a switching power supply. Specifically, the fluorescence quantitative PCR analyzer is an existing mature application. The control module, temperature control module, photoelectric module, power module and other related components are not shown in the embodiment. Their specific structure can refer to the existing technology. Their installation position can be adapted according to actual needs. Here, no further details are given.

[0050] In this embodiment, when amplifying or detecting DNA, DNA templates, primers, enzymes, and other raw materials are added to the corresponding PCR tubes according to the detection requirements. When the PCR tubes have different heights, PCR tubes of different heights are placed inside multiple loading cylinders 12. The heat-conducting cover 13 is rotated so that it rotates closer to the loading cylinder 12, changing the device from an open state to a closed state. When multiple heat-conducting sleeves 21 come into contact with PCR tubes of different heights, the PCR tubes push the heat-conducting sleeves 21 upwards, compressing the elastic element 22 through the heat-conducting sleeves 21. PCR tubes of different heights can push the heat-conducting sleeves 21 they are in contact with to different heights. Because the elastic element 22 is in a compressed state, under the drive of the elastic element 22, the heat-conducting sleeves 21 can always maintain contact with the PC. The cap at the top of the R tube fits tightly. When the device is started, the raw materials in the PCR tube are circulated and heated by the temperature control module. The temperature control module also causes the conductive heating film 16 to release heat, which heats the heat-conducting plate 15. The heat-conducting plate 15 absorbs the heat and transfers it to the heat-conducting sleeve 21, causing the heat-conducting sleeve 21 to heat up and preventing condensation of the reaction solution in the PCR tube. With the above scheme, each PCR tube has an independent heat-conducting sleeve 21, and the cap of the PCR tube is pressed together by the heat-conducting sleeve 21 to maintain the airtightness of the PCR tube. While maintaining the high capacity advantage of a single tank, the device can accommodate PCR tubes of different heights, meeting diverse monitoring needs and improving the applicability of the device. This fundamentally solves the technical conflict between the equipment structure and the diversity of test samples.

[0051] It should be noted that when amplifying DNA (cycle heating), 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-conducting sleeve 21 is 104-108°C to prevent the reaction solution (raw material) in the PCR tube from evaporating and from condensing with the cap on the PCR tube.

[0052] Secondly, please refer to it again. Figures 3 to 6As shown, the adjustment assembly 30 includes a guide sleeve 31, a drive motor 32, a threaded rod 33, and a base 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 housing 13 and located at the upper end of the guide sleeve 31, and the output end of the drive motor 32 extends into the interior of the guide sleeve 31. The threaded rod 33 is fixed to the output end of the drive motor 32, and the lower end of the outer side of the threaded rod 33 is located inside the elastic element 22. The threaded rod 33 and the guide sleeve 31 are rotatably connected through a sealed bearing. The base plate 34 is threaded to the outer side of the threaded rod 33. The base plate 34 and the elastic element 22 are adapted to each other, and the base plate 34 and the guide sleeve 31 are slidably connected. After the drive motor 32 is running, the base plate 34 can adjust the compression of the elastic element 22.

[0053] Here, the guide sleeve 31 and the control module are electrically connected by wires. In this embodiment, the guide sleeve 31 is preferably a servo motor.

[0054] In this embodiment, when amplifying DNA, PCR tubes of different heights are loaded into the loading cylinder 12 according to the detection requirements. The heat-conducting cap 13 is rotated to change the device from an open to a closed state. After the PCR tubes of different heights and multiple heat-conducting sleeves 21 are attached one by one, the PCR tubes push the heat-conducting sleeves 21 upwards, compressing the elastic element 22 through the heat-conducting sleeves 21. Due to the different heights of the PCR tubes, the amount of compression of the elastic element 22 by the heat-conducting sleeves 21 is also different (the compression of the elastic element 22 at the upper end of the relatively shorter PCR tube is smaller, and the compression of the elastic element 22 at the upper end of the relatively taller PCR tube is larger). When the elastic element 2... When the compression of element 2 is too large, the elastic element 22 applied to the corresponding heat-conducting sleeve 21 and PCR tube will also have a large rebound force, which may easily cause the PCR tube to deform and crack. The corresponding drive motor 32 is started. Through the fixed connection between the drive motor 32 and the threaded rod 33, the drive motor 32 drives the threaded rod 33 to rotate. Through the fixed connection between the threaded rod 33 and the base plate 34 and the sliding connection between the base plate 34 and the guide sleeve 31, the threaded rod 33 drives the base plate 34 to move upward. After the base plate 34 moves upward, the elastic element 22 with a large compression can be released appropriately, thereby reducing the rebound force applied to the PCR tube and preventing the PCR tube from being deformed or cracked due to a large rebound force.

[0055] Secondly, please refer to the following as well. Figures 4 to 6 As shown, a pressure sensing element 35 is fixed at the lower end of the base plate 34, and the sensing end of the pressure sensing element 35 is in contact with the upper end of the elastic element 22.

[0056] Here, the pressure sensing element 35 and the control module are electrically connected by wires. A touch display component is provided at the upper end of the analyzer body 10 away from the hot cover housing 13. 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.

[0057] Furthermore, since the elastic element 22 is always in a compressed state, and the upper and lower ends of the elastic element 22 are in contact with the sensing end of the pressure sensing element 35 and the heat-conducting sleeve 21 respectively, during the detection process, the bottom of the heat-conducting sleeve 21 is tightly in contact with the top cap of the PCR tube. The rebound force of 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 of the elastic element 22 on the PCR tube. During this process, the rebound force of the compressed elastic element 22 on the sensing end of the pressure sensing element 35, the rebound force on the heat-conducting sleeve 21, and the pressing force on the PCR tube are all equal.

[0058] In this embodiment, during DNA amplification, the compression of the elastic element 22 corresponding to each PCR tube with different heights is not the same, and the pressure applied by each elastic element 22 to the PCR tube is also not the same. The pressure sensing element 35 senses the rebound force of the corresponding elastic element 22. When the compression of the elastic element 22 is too large and the rebound force is greater than the preset pressure target value, the drive motor 32 is activated. The drive motor 32 drives the base plate 34 and the pressure sensing element 35 to move upward, so that the elastic element 22 with the large compression is released until the rebound force of the elastic element 22 is equal to the pressure target value; conversely, when the compression of the elastic element 22 is too large, the rebound force is greater than the preset pressure target value. If the compression amount of the elastic element 22 is too small and the rebound force is less than the preset pressure target value, the drive motor 32 is started. The drive motor 32 drives the base plate 34 and the pressure sensing element 35 to move downward, so that the elastic element 22 with the small compression amount is further compressed until the rebound force of the 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 of different heights, ensuring that the pressure applied to each PCR tube is consistent. This avoids the problems of poor PCR tube sealing and reaction liquid evaporation caused by insufficient pressure, and also prevents PCR tube deformation or cracking caused by excessive pressure.

[0059] Secondly, please refer to it again. Figure 5 and Figure 6 An anti-detachment plate 36 is fixed at the lower end of the threaded rod 33 and inside the elastic element 22.

[0060] It should be noted that the pressure sensing element 35 is preferably an annular pressure sensor. The annular pressure sensor has a through hole coaxially opened inside. 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.

[0061] In this embodiment, when the drive motor 32 is started to move the base plate 34 and the pressure sensing element 35 in the vertical direction, the anti-detachment plate 36 can prevent the threaded rod 33 from separating from the base plate 34 and the pressure sensing element 35.

[0062] Please refer to it again. Figure 4 and Figure 6 The guide sleeve 31 has a guide boss 37 inside, the bottom plate 34 has a second guide groove 38 on the outside, and the heat-conducting sleeve 21 has a first guide groove 23 on the outside. The bottom plate 34 and the second guide groove 38, as well as the bottom plate 34 and the first guide groove 23, are all compatible with each other. The guide sleeve 31 and the bottom plate 34 are slidably connected by the cooperation of the guide boss 37 and the second guide groove 38, and the guide sleeve 31 and the heat-conducting sleeve 21 are slidably connected by the cooperation of the guide boss 37 and the first guide groove 23.

[0063] In this embodiment, when adjusting the compression of the elastic element 22, the drive motor 32 is started to adjust the height of the base plate 34 and the pressure sensing element 35. Through the coordinated work of the guide boss 37 and the second guide groove 38, the guide sleeve 31 is able to guide the base plate 34 during its movement, preventing the base plate 34 from rotating.

[0064] Please refer to it again. Figures 4 to 6 The guide sleeve 31 has an clearance groove 39 on its outer side, and the clearance groove 39 is compatible with the pressure sensing element 35.

[0065] In this embodiment, since the pressure sensing element 35 and the control module are electrically connected by wires, when the pressure sensing element 35 moves, the connected wires will also move synchronously accordingly. The clearance groove 39 is designed to ensure that the wires can move smoothly and synchronously.

[0066] Please refer to it again. Figure 3 The upper end of the heat cover housing 13 is provided with a second groove 17, and a protective plate 18 is fixed to the upper end of the second groove 17. The drive motor 32 is fixed inside the second groove 17, and the second groove 17 and the first groove 14 are not connected.

[0067] In this embodiment, during the DNA amplification process, the conductive heating film 16 releases heat, which causes the internal temperature of the first groove 14 to rise synchronously. When the DNA is denatured, the internal temperature of the first groove 14 reaches 104-108°C. By setting an independent second groove 17 to fix the drive motor 32, the drive motor 32 can be prevented 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.

[0068] Here, a heat insulation layer (e.g., a ceramic fiber layer) is also fitted at the upper end of the first groove 14.

[0069] It should be noted that a shaft through hole is provided between the first groove 14 and the second groove 17. The output end of the drive motor 32 extends into the guide sleeve 31 through the shaft through hole. Since the output end of the drive motor 32 is inserted into the shaft through hole, and the top of the guide sleeve 31 can also prevent air convection from occurring between the first groove 14 and the second groove 17, the first groove 14 and the second groove 17 are described as not connected in this application.

[0070] 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, chromium silicon alloy steel, nickel titanium alloy or other materials that can be used to make springs and can be used for a long time in high temperature environments above 120°C. In this embodiment, the material of the elastic element 22 is preferably 302 stainless steel.

[0071] In this embodiment, when the DNA is denatured, the temperature inside the first groove 14 is as high as 104 to 108°C. With the above-mentioned scheme, the elastic element 22 can work stably at high temperature, avoiding the failure of the elastic element 22 due to high temperature.

[0072] Example 2

[0073] A detection method for a Gram-positive bacterial pathogen PCR detection device, applicable to any one of the Gram-positive bacterial pathogen PCR detection devices in Example 1, includes the following steps:

[0074] St1: Add the DNA template, primers, enzymes and other raw materials to the corresponding PCR tubes, and then load the multiple PCR tubes into the multiple loading tubes 12 respectively;

[0075] St2: Set the target pressure value, change the device from the open state to the closed state, sense the rebound force 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 rebound force of the elastic element 22 so that the pressing force applied by each elastic element 22 to the corresponding PRC pipe is the same as the target pressure value.

[0076] St3: Start the device to amplify or detect DNA.

[0077] Furthermore, this device integrates real-time quantitative PCR (qPCR) technology during DNA amplification, enabling precise quantitative analysis of nucleic acids through dynamic monitoring of fluorescence signals. Specifically, the photoelectric module incorporates a multi-channel fluorescence detection system, employing either the SYBR Green I fluorescent dye method or the TaqMan probe method to track amplification products in real time. Upon device startup, the temperature control module drives the conductive heating film 16 and the heat-conducting plate 15 to cycle according to a preset program (denaturation: 94-98℃, annealing: 55-65℃, extension: 72℃). Simultaneously, the photoelectric module periodically excites the fluorescence signal within the PCR tube using an excitation light source (such as an LED). For the SYBR Green I method, fluorescence intensity is directly proportional to the double-stranded DNA content; the signal is acquired by the photoelectric converter and transmitted to the control module. If the TaqMan probe method is used, the probe is cleaved by Taq enzyme during amplification, separating the fluorescent group from the quenching group, and the fluorescence signal intensifies as the product accumulates. The software system built into the control module plots fluorescence amplification curves in real time. By comparing the threshold cycle number (Ct value) with the standard curve, it automatically calculates the initial template concentration. This process is visualized through a touch display component, allowing users to directly obtain quantitative results of pathogen nucleic acids. In addition, the device supports multi-fluorescence channel detection (such as FAM, HEX, VIC), and can simultaneously analyze multiple target genes, meeting complex detection needs such as Gram-positive bacteria drug resistance gene typing.

[0078] In one specific embodiment, this embodiment provides a method for detecting Gram-positive bacterial pathogens based on real-time quantitative PCR (qPCR) technology, the specific steps of which are as follows:

[0079] Step 1: Sample preparation and reaction system configuration

[0080] Extract DNA or RNA from the sample to be tested and synthesize cDNA using a reverse transcription kit (for RNA samples). Prepare the PCR reaction solution according to the specified proportions, containing the following components:

[0081] 10×PCR buffer (containing Mg) 2+ )

[0082] dNTP mixture (200 μm each)

[0083] Forward / reverse primers (0.2 μm each)

[0084] SYBR Green I fluorescent dye or TaqMan probe (0.1 μm)

[0085] Taq DNA polymerase (1 U / μL)

[0086] Template DNA / cDNA (2-5 μL)

[0087] Step 2: Loading and Parameter Settings

[0088] Dispense the prepared reaction solution into PCR tubes and load them into the loading tube 12 of the analyzer body 10. Set the qPCR program via the touch display component:

[0089] Pre-denaturation: 95℃, 3 min

[0090] Cyclic amplification (40 cycles): denaturation at 95°C for 15 seconds → annealing at 60°C for 30 seconds → extension at 72°C for 30 seconds;

[0091] Melting curve analysis: 95℃, 15s → 60℃, 1min → 95℃, 15s (continuous monitoring of fluorescence signal).

[0092] Step 3: Real-time fluorescence detection and data analysis

[0093] After the device is started, the photoelectric module acquires fluorescence signals from 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 a standard curve to quantify the copy number of the target gene in the sample. If melting curve analysis is used, the specificity of the amplified product can be verified through the characteristic melting temperature (Tm value). This embodiment achieves high sensitivity and high specificity detection of Gram-positive bacterial pathogens by integrating qPCR technology. The multi-channel fluorescence synchronous acquisition function supports multiplex PCR detection, which can simultaneously analyze multiple drug resistance genes (such as mecA and vanA), significantly improving detection efficiency. Combined with the adaptive compression component in Example 1, the sealing of PCR tubes of different sizes is ensured, avoiding deviations in fluorescence signals due to evaporation or condensation.

[0094] The working principle of the present invention is:

[0095] The pressure target value is set via the touch display component. When amplifying or detecting DNA, raw materials such as DNA templates, primers, and enzymes are added to the corresponding PCR tubes according to the detection requirements. PCR tubes of different heights are then loaded into the interiors of multiple loading cylinders 12. Rotating the heat-conducting cap 13 changes the device from an open to a closed state. When multiple heat-conducting sleeves 21 come into contact with PCR tubes of different heights, the PCR tubes push the heat-conducting sleeves 21 upwards, compressing the elastic element 22. PCR tubes of different heights can push the heat-conducting sleeves 21 they are in contact with to different heights. Driven by the compressed elastic element 22, the heat-conducting sleeve 21 can always maintain a tight seal with the top cap of the PCR tube. At the same time, the compression amount of each elastic element 22 is not exactly the same. The pressure sensing element 35 senses the corresponding... If the rebound force of the elastic element 22 is not the same as the target pressure value, the drive motor 32 is activated. The drive motor 32 drives the base plate 34 and the pressure sensing element 35 to move vertically, thereby automatically adjusting the compression amount of the elastic element 22 until the rebound force of the elastic element 22 is the same as the target pressure value. Then the device is started to amplify or detect DNA. This allows the device to load PCR tubes of different heights while maintaining the high capacity advantage of a single tank, meeting diverse monitoring needs and improving the applicability of the device. It can also independently adjust the compression amount of each elastic element 22 for PCR tubes of different heights to ensure that the pressure applied to each PCR tube is consistent. This avoids problems such as poor sealing of PCR tubes and evaporation of reaction solution due to insufficient pressure, and also prevents deformation or cracking of PCR tubes due to excessive pressure.

[0096] The above are merely preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention. Structures, devices, and operating methods not specifically described or explained in this invention are implemented according to conventional methods in the art unless otherwise specified or limited.

Claims

1. A PCR detection device for Gram-positive bacterial pathogens, characterized in that: The analyzer includes an analyzer body (10), the analyzer body (10) having a loading groove (11) inside, the loading groove (11) having multiple loading cylinders (12) inside, one end of the analyzer body (10) being rotatably connected to a heat cover shell (13), the lower end of the heat cover shell (13) having a first groove (14), the first groove (14) having a heat-conducting plate (15) and a conductive heating film (16) fixed inside, and the conductive heating film (16) being located above the heat-conducting plate (15), and also includes: Multiple pressing components (20) are assembled inside the heat-conducting plate (15), and there is a one-to-one correspondence between the multiple pressing components (20) and the multiple loading cylinders (12). Each pressing component (20) includes a heat-conducting sleeve (21) and an elastic element (22). The heat-conducting sleeve (21) is slidably connected to the inside of the heat-conducting plate (15), and the elastic element (22) is assembled inside the heat-conducting sleeve (21). Multiple adjustment components (30) are assembled inside the heat cover housing (13), and there is a one-to-one correspondence between the multiple adjustment components (30) and the multiple pressing components (20). Each adjustment component (30) includes a guide sleeve (31), a drive motor (32), a threaded rod (33), and a base 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 to the heat cover housing (13). 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 base plate (34) is threaded to the outside of the threaded rod (33). The base plate (34) and the elastic element (22) are adapted to each other. The base plate (34) and the guide sleeve (31) are slidably connected. The lower end of the base plate (34) is fixed with a pressure sensing element (35). The pressure sensing element (35) and the elastic element (22) are in contact. The elastic element (22) is always in a compressed state, and the elastic element (22) enables multiple heat-conducting sleeves (21) to fit tightly against the top of PCR tubes of different heights.

2. The PCR detection device for Gram-positive bacterial pathogens according to claim 1, characterized in that: The lower end of the threaded rod (33) is fixed with an anti-detachment plate (36).

3. The PCR detection device for Gram-positive bacterial pathogens according to claim 1, characterized in that: The guide sleeve (31) is provided with a guide boss (37) inside, the bottom plate (34) is provided with a second guide groove (38) on the outside, the heat-conducting sleeve (21) is provided with a first guide groove (23) on the outside, and the bottom plate (34) and the second guide groove (38) and the bottom plate (34) and the first guide groove (23) are all compatible with each other.

4. The PCR detection device for Gram-positive bacterial pathogens according to claim 1, characterized in that: The guide sleeve (31) has an clearance groove (39) on its outer side, and the clearance groove (39) is compatible with the pressure sensing element (35).

5. The PCR detection device for Gram-positive bacterial pathogens according to claim 1, characterized in that: The upper end of the heat cover housing (13) is provided with a second groove (17), and a protective plate (18) is fixed at the upper end of the second groove (17). The drive motor (32) is fixed inside the second groove (17).

6. The PCR detection device for Gram-positive bacterial pathogens according to claim 1, characterized in that: The elastic element (22) is a helical spring.

7. The PCR detection device for Gram-positive bacterial pathogens according to claim 6, characterized in that: The elastic element (22) is made of any one of the following materials: 302 stainless steel, chromium silicon alloy steel, or nickel titanium alloy.

8. A detection method for Gram-positive bacterial pathogens using a PCR detection device for non-therapeutic and non-diagnostic purposes, applicable to the Gram-positive bacterial pathogen PCR detection device according to any one of claims 1 to 7, characterized in that: Includes the following steps: St1: Add the raw materials to the corresponding PCR tube, and then load the PCR tube into the loading tube (12); St2: Set the target pressure value, change the device from the open state to the closed state, and adjust the compression of each elastic element (22) by adjusting the adjustment component (30); St3: Start the device to amplify the DNA.

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