A nucleic acid detection analyzer and its detection method

Abstract

The present invention relates to nucleic acid detection, and specifically to a nucleic acid detection analyzer and a detection method thereof. The nucleic acid detection analyzer includes a frame, a temperature control device mounted on the frame, a fluorescence detection device, a hot cover device, and a reagent tube; the frame includes a sample loading platform, and the sample loading platform is provided with an insertion hole; the temperature control device and the fluorescence detection device are mounted between the base plate and the sample loading platform; the hot cover device is mounted on the frame; the reagent tube is mounted on the insertion hole and covered by the hot cover device; the position of the reagent tube is adapted to the position of the temperature control device; and the detection position of the fluorescence detection device is directly opposite to the reagent tube. The present invention has multiple scenarios, low operating requirements, and a price suitable for promotion.

Description

Technical Field

[0001] The present invention relates to nucleic acid detection, and in particular to a nucleic acid detection analyzer and a detection method thereof. Background Art

[0002] Nucleic acid detection technology is a molecular biology detection method that requires first extracting nucleic acid fragments (DNA or RNA) from the sample, then replicating and amplifying the nucleic acid fragments through polymerase chain reaction, and finally detecting the nucleic acid to determine the specific type or source of the nucleic acid in the sample.

[0003] Nucleic acid testing is simple and widely used, requiring only a small sample size to produce accurate results. PCR testing first requires sampling, typically using a swab to collect the sample into a tube. The sample is then extracted from the tube into a reagent kit, and finally placed into the instrument for reaction. During the nucleic acid extraction and amplification process, the first and second reagents are added to the sample in separate steps and mixed.

[0004] Existing nucleic acid testing protocols generally require two instruments: a nucleic acid extraction instrument is used to extract the nucleic acid, and the extracted nucleic acid is manually transferred to a reagent kit. The kit is then installed in a corresponding nucleic acid detection analyzer (such as a QPCR instrument) for testing. This protocol uses multiple devices and has many manual steps, requiring professional operators and a specific environment to prevent cross-contamination. The entire testing process is also relatively time-consuming.

[0005] Currently, there are nucleic acid extraction workstations that connect nucleic acid extraction equipment and detection equipment through a mechanical wall to achieve full automation of the nucleic acid transfer process. However, nucleic acid extraction workstations are complex in structure, bulky, and expensive, and require professional operation.

[0006] With the increasing demand for nucleic acid testing and the popularization and promotion of nucleic acid testing technology, the market urgently needs to launch a nucleic acid testing analyzer with multiple application scenarios, low operating requirements and a price suitable for promotion. Summary of the Invention

[0007] The present invention provides a nucleic acid detection analyzer and a detection method thereof, so as to solve the problems of existing instruments such as complex operation, limited application scenarios, complex structure and high cost.

[0008] In order to achieve the above purpose, the present invention adopts the following technical means:

[0009] A nucleic acid detection analyzer includes a frame, a temperature control device installed on the frame, a fluorescence detection device, a hot cover device and a reagent tube.

[0010] It includes a bottom plate and a sample loading platform installed on the bottom plate, wherein the sample loading platform is provided with an insertion hole;

[0011] The temperature control device and the fluorescence detection device are installed between the bottom plate and the sample loading platform;

[0012] The thermal cover device is installed on the frame;

[0013] The reagent tube is installed on the insertion hole and covered by the hot cover device;

[0014] The position of the reagent tube is adapted to the position of the temperature control device (the adaptation mentioned here means that after the reagent tube is installed, one end of the reagent tube is located in the temperature control device, and the temperature control device is capable of heating the reagent tube); the detection position of the fluorescence detection device is facing the reagent tube (the detection position mentioned here is facing the reagent tube, and the fluorescence detection device is capable of detecting particles in the reagent tube); the detection position of the fluorescence detection device is facing the reagent tube.

[0015] One end of the test tube covered by the hot cover is located above the sample loading platform, and the other end is located below the sample loading platform. The fluorescence detection device is located below the reagent tube. The temperature control device includes a movable platform for heating the lower end of the reagent tube, and the movable platform is located between the sample loading platform and the fluorescence detection device.

[0016] The working principle of the present invention is:

[0017] Add the sample to the reagent tube to mix the sample with the first reagent, open the hot cover device, put the reagent tube into the insertion hole and cover the hot cover device,

[0018] Heat the hot cover device,

[0019] The temperature control device heats and incubates the reaction site of the reagent tube, releasing the nucleic acid in the sample into the reagent tube.

[0020] Add the second reagent in the reagent storage tube to the reagent tube and mix it with the sample.

[0021] Controlling the temperature of the reaction site of the reagent tube using a temperature control device so that the sample reagent mixture in the reagent tube reaches a first temperature and the nucleic acid is melted at the first temperature;

[0022] controlling the temperature of the reaction portion of the reagent tube using a temperature control device so that the sample-reagent mixture in the reagent tube reaches a second temperature, and the nucleic acid replicates at the second temperature;

[0023] Controlling the temperature of the reaction portion of the reagent tube using a temperature control device so that the temperature of the sample reagent mixture in the reagent tube cycles between the first temperature and the second temperature until the amplification of the nucleic acid is completed;

[0024] The fluorescence detection device performs optical detection on the nucleic acid in the reagent tube.

[0025] The number of the temperature control device, the fluorescence detection device, the hot cover device and the reagent tubes is at least two, and the temperature control device, the fluorescence detection device, the hot cover device and the reagent tubes are matched one by one.

[0026] The frame further comprises a bottom plate, a support frame and a shell mounted on the bottom plate. The sample loading platform is mounted on the support frame, and the hot cover device is mounted on the sample loading platform.

[0027] The temperature control device includes a drive component, a guide component, and a first heating component; the first heating component includes a mobile platform, and the mobile platform has more than three heating grooves for heating. The end of the reagent tube is located in a heating groove. The mobile platform moves along the guide component under the drive of the drive component, so that the end of the reagent tube is located in different heating grooves.

[0028] The driving assembly includes a temperature-controlled driving motor installed on the frame, a bearing seat, a screw with one end connected to the temperature-controlled driving motor and the other end connected to the bearing seat, and a screw nut connected to the screw; the guide assembly includes two guide rails installed on the frame; the movable platform of the first heating assembly is installed on the screw nut and is equipped with a slider that is slidably connected to the guide rail.

[0029] There are four heating tanks, which are arranged parallel to the guide rails. Each heating tank is equipped with a ceramic heating plate and a heat-conducting copper plate that is in close contact with the ceramic heating plate. There are two heat-conducting copper plates in each heating tank, and a gap is provided between the two heat-conducting copper plates for the reagent tube to pass through. The heat-conducting copper plates are in contact with the outer wall of the reagent tube.

[0030] The temperatures in the four heating tanks include a first target temperature and a second target temperature for polymerase chain reaction, a first transition temperature and a second transition temperature for transition, and the values of the first transition temperature, the first target temperature, the second target temperature, and the second transition temperature decrease in sequence.

[0031] The mobile platform is made of heat-insulating material; the mobile platform is composed of two interconnected heat-insulating boards; the temperature-controlled drive motor is a stepping motor; a trigger is installed on the mobile platform, and an induction switch is installed on the frame, and the positions of the trigger and the induction switch are adapted; or a trigger is installed on the frame, and an induction switch is installed on the mobile platform, and the positions of the trigger and the induction switch are adapted (the adaptation mentioned here means that the relative movement of the trigger and the induction switch can control the induction switch to be triggered).

[0032] The heat cover device includes a cover shell and a second heating assembly installed on the cover shell; the second heating assembly includes a heat conducting plate and a heating element in contact with the heat conducting plate, the lower surface of the heat conducting plate has a downwardly protruding annular boss, the shape of the annular boss is adapted to the end of the reagent tube (the adaptation mentioned here means that the annular boss can surround the end of the reagent tube so that the heating space covers the end of the reagent tube), and the inner wall of the annular boss and the lower surface of the heat conducting plate form a heating space for heating the end of the reagent tube.

[0033] The heat cover device also includes a guide structure installed on the cover shell, the guide structure includes a guide plate, a guide shaft installed on the lower surface of the guide plate, and a first elastic member, and the first elastic member is located between the heat conducting plate and the guide plate; a guide hole is opened on the heat conducting plate, and the guide hole is cooperatively connected with the guide shaft.

[0034] A first push rod through hole is provided on the heat conducting plate of the second heating assembly, a second push rod through hole and a waist-shaped hole are provided on the guide plate, and the end of the push rod structure passes through the first push rod through hole and the second push rod through hole and is located in the heating space.

[0035] The cover shell includes an upper cover shell and a lower cover shell connected to the upper cover shell. The upper cover shell and the lower cover shell are both hollow structures. The upper end of the upper cover shell is sealed, and the inner wall of the lower cover shell has a first mounting step and a second mounting step; the guide plate of the guide structure is installed on the first step, and the heat conduction plate of the second heating assembly is located on the second step.

[0036] The hot cover device also includes a push rod structure, which includes a push rod mounting frame installed on the cover shell, a push rod driving member installed on the push rod mounting frame, and a push rod slidably arranged on the push rod mounting frame and cooperatively connected to the push rod driving member. The driving member can drive the end of the push rod into the heating space.

[0037] The ejector drive is a linear motor, and the ejector mounting bracket is equipped with a linear bearing, on which the ejector is mounted. A third groove is defined in the middle of the ejector, which engages with the slider of the linear motor. The ejector end has an ejector boss. (The ejector boss is used to pierce the sealing membrane at a location corresponding to the position of the reagent storage tube and engages with the first groove at the top of the reagent storage tube to ensure that the reagent storage tube is subjected to a vertical downward force, thereby pushing the reagent storage tube downward.)

[0038] The reagent tube includes a tube cover and a reaction tube, the tube cover is provided with a sample addition hole and a reagent addition hole; a reagent storage tube is installed in the reagent addition hole, the reagent storage tube is filled with a second reagent, and the tube cover is provided with a sealing film for sealing the sample addition hole and the reagent addition hole; the reaction tube has a reaction cavity; the tube cover and the reaction tube are connected in a coordinated manner so that the reaction cavity is in a sealed state.

[0039] The reagent storage tube is movably installed in the reagent adding hole of the tube cover.

[0040] The reagent storage tube is tubular, with one end being closed and the other end being open; the open end of the reagent storage tube has an oblique edge, and the closed end is provided with a first groove.

[0041] The reagent storage tube also has a gravity ball, the second reagent is a freeze-dried ball, and the gravity ball is placed above the freeze-dried ball.

[0042] The gravity ball is a solid with a smooth surface and is made of polymer material.

[0043] The reaction tube contains a first reagent, and the lower half of the reaction tube has a transparent detection window; the detection window is located at the bottom of the reaction tube.

[0044] Both the openings at both ends of the sample addition hole and the openings at both ends of the reagent addition hole are provided with sealing films, and both the sample addition hole and the reagent hole are circular through holes.

[0045] A reagent tube positioning groove is provided on the sample loading platform, the cross-section of the tube cover is circular or square, and a first protrusion is provided on the outer wall of the tube cover, which is adapted to the reagent tube positioning groove (the adaptation mentioned here means that the first protrusion can cooperate with the reagent tube positioning groove and can position the reagent tube); a rubber sealing plug is fixed in the sample loading hole; the lower half of the reaction tube is flat.

[0046] The cross-section of the upper half of the reaction tube is circular or rectangular, and the upper half of the reaction tube is connected to the tube cover. A sealing film is provided at the opening of the reaction tube to seal one end of the sample addition hole and the reagent addition hole, and a sealing film is provided on the upper surface of the tube cover to seal the other end of the sample addition hole and the reagent addition hole. The circular or rectangular shape is easy to process and convenient to take and put. In addition, the circular tube wall can ensure uniform thickness, which is conducive to ensuring that the temperature inside the reaction tube is stable at the target value through heating. This structure is wider at the top, which can ensure that the reagent and sample are fully in contact and react, and ensure the dissolution of the freeze-dried ball. The narrower lower structure ensures the heat conduction speed and PCR amplification speed.

[0047] A detection method of a nucleic acid detection analyzer, characterized in that: the nucleic acid detector includes a frame, a temperature control device installed on the frame, a fluorescence detection device, a hot cover device and a reagent tube,

[0048] The frame includes a bottom plate and a support frame mounted on the bottom plate, a shell, and a sample loading platform mounted on the support frame, wherein the sample loading platform is provided with an insertion hole;

[0049] The temperature control device and the fluorescence detection device are installed between the bottom plate and the sample loading platform;

[0050] The thermal cover device is installed on the frame;

[0051] The reagent tube is installed on the insertion hole and covered by the hot cover device, the reagent tube contains a first reagent and a reagent storage tube separated from the first reagent, and the reagent storage tube contains a second reagent;

[0052] The position of the reagent tube is adapted to the position of the temperature control device; the detection position of the fluorescence detection device is directly opposite to the reagent tube;

[0053] The detection method comprises:

[0054] Step 1: Add the sample to the reagent tube to mix the sample with the first reagent, open the hot cover device, put the reagent tube into the insertion hole and cover the hot cover device,

[0055] Step 2: The temperature control device heats and incubates the reaction area of the reagent tube to release the nucleic acid in the sample into the reagent tube.

[0056] Step 3: Add the second reagent in the reagent storage tube to the reagent tube and mix it with the sample.

[0057] Step 4: Using a temperature control device to control the temperature of the reaction site of the reagent tube, so that the sample and reagent mixture in the reagent tube reaches a first temperature, and the nucleic acid is melted at the first temperature;

[0058] Step 5: Using a temperature control device to control the temperature of the reaction portion of the reagent tube, so that the sample and reagent mixture in the reagent tube reaches a second temperature, and the nucleic acid replicates at the second temperature;

[0059] Step 6: Using a temperature control device to control the temperature of the reaction portion of the reagent tube, so that the temperature of the sample reagent mixture in the reagent tube cycles between the first temperature and the second temperature until the amplification of the nucleic acid is completed;

[0060] Step 7: The fluorescence detection device performs optical detection on the nucleic acid in the reagent tube.

[0061] Furthermore, the method further includes step A: heating the thermal cover device; step A is performed simultaneously with step 6 or before step 6. Heating the thermal cover can prevent uneven heating of the reagent chamber and avoid condensation. It is understood that heating the thermal cover simultaneously with step 6 or before step 6 can achieve this effect, and the best effect is achieved when the thermal cover is heated in step 4 or before step 4.

[0062] The beneficial effects of the present invention are:

[0063] The present invention directly adds the sample into a dedicated reagent tube, and places the reagent tube in a specific environment through a hot cover device. The hot cover device and a temperature control device create a reaction environment for the reagent tube, and the second reagent can be added without opening the hot cover, thereby completing the entire polymerase chain reaction process.

[0064] Since the entire polymerase chain reaction process is fully enclosed and pollution-free, does not require a dedicated laboratory environment, and only requires sample addition operations, without the need for professional operators, the present invention can be applied to various scenarios and has a wide range of applications.

[0065] The reaction process of the present invention is also relatively simple, the reaction time is short, the test results can be obtained quickly, and the device of the present invention is compact and can be carried in a bag. These advantages make the present invention suitable for bedside testing and further expand the scope of application of the present invention.

[0066] Since the various parts of the present invention are highly integrated and independent of each other, that is, a reagent tube, a hot cover device, a temperature control device and an optical detection device can be used together on the rack to perform a complete detection process; therefore, the structure of the present invention can be expanded according to the detection requirements and transformed into a multi-channel nucleic acid detection analyzer, such as Figure 1 As shown, two reagent tubes, two thermal cover devices, two temperature control devices, and two optical detection devices are installed in a rack to form a dual-throughput (i.e., simultaneous detection of two samples) nucleic acid detection analyzer. Similarly, utilizing the principles of the present invention, the number of samples that can be simultaneously detected by the nucleic acid detection analyzer can be increased, such as to three, four, or eight channels, allowing the present invention to flexibly adjust the number of detection channels according to different needs. BRIEF DESCRIPTION OF THE DRAWINGS

[0067] Figure 1 It is a schematic diagram of the external structure of the present invention;

[0068] Figure 2 It is a schematic diagram of the positional relationship of various parts of the present invention;

[0069] Figure 3 It is a schematic diagram of the internal structure of the present invention;

[0070] Figure 4 It is a structural diagram of the temperature control device;

[0071] Figure 5 It is a structural diagram of the mobile platform of the temperature control device;

[0072] Figure 6 1 is a schematic diagram of the external structure of the thermal cover device;

[0073] Figure 7 It is a structural schematic diagram of the cover shell of the thermal cover device;

[0074] Figure 8 is a schematic diagram of the internal structure of the hot cover device;

[0075] Figure 9 It is a schematic diagram of the structure of the heat conducting plate of the heat cover device;

[0076] Figure 10 It is a schematic diagram of the structure of the reagent tube;

[0077] Figure 11 yes Figure 10 sectional view of .

[0078] The numbers in the figure represent:

[0079] 11-base plate, 12-sample loading platform, 13-support frame, 14-housing;

[0080] 2-temperature control device, 21-temperature control drive motor, 22-bearing seat, 23-screw, 24-screw nut, 25-guide rail, 26-slider, 27-moving platform, 28-ceramic heating plate, 29-thermal copper plate, 210-trigger, 211-induction switch;

[0081] 3-fluorescence detection device;

[0082] 4-heat cover device, 41-cover shell, 411-upper cover shell, 412-lower cover shell, 413-first mounting step, 414-second mounting step, 42-heat conducting plate, 421-annular boss, 422-sensor mounting groove, 43-guide plate, 431-first elastic member, 432-waist-shaped hole, 441-rod mounting bracket, 442-linear motor, 443-rod, 444-linear bearing;

[0083] 5-reagent tube, 51-tube cover, 52-reaction tube, 53-reaction chamber, 54-reagent storage tube, 55-rubber sealing plug, 56-first protrusion, 57-gravity ball, 58-lyophilization ball, 59-oblique edge, 510-first groove;

[0084] 6-Display screen. Specific embodiments

[0085] The present invention provides a method to solve the problems of complex operation, limited application scenarios, complex structure and high cost of existing instruments.

[0086] like Figure 1 、 Figure 2 、 Figure 3As shown, a nucleic acid detection analyzer includes a frame, a temperature control device 2 installed on the frame, a fluorescence detection device 3, a hot cover device 4 and a reagent tube 5, the frame includes a base plate 11 and a support frame 13 installed on the base plate, a shell 14, and a sample loading table 12 installed on the support frame 13, and the sample loading table 12 is provided with an insertion hole; the temperature control device 2 and the fluorescence detection device 3 are installed between the base plate 11 and the sample loading table 12; the hot cover device 4 is installed on the frame; the reagent tube 5 is installed on the insertion hole and covered by the hot cover device 4; the position of the reagent tube 5 is adapted to the position of the temperature control device 2 (the adaptation mentioned here means that after the reagent tube 5 is installed, one end of the reagent tube is located in the temperature control device, and the temperature control device can control the temperature in the reagent tube); the detection position of the fluorescence detection device 3 is opposite to the reagent tube 5.

[0087] In this embodiment, the hot cover device 4 is installed on the sample loading platform 12 .

[0088] One end of the reagent tube covered by the hot cover is located above the sample loading platform, and the other end is located below the sample loading platform. The fluorescence detection device is located below the reagent tube. The temperature control device includes a movable platform for heating the lower end of the reagent tube. The movable platform is located between the sample loading platform and the fluorescence detection device.

[0089] A detection method of a nucleic acid detection analyzer, characterized in that: the nucleic acid detector includes a frame, a temperature control device 2 installed on the frame, a fluorescence detection device 3, a hot cover device 4 and a reagent tube 5,

[0090] The frame includes a bottom plate 11 and a support frame 13 mounted on the bottom plate, a housing 14, and a sample loading platform 12 mounted on the support frame 13. The sample loading platform 12 is provided with an insertion hole.

[0091] The temperature control device 2 and the fluorescence detection device 3 are installed between the base plate 11 and the sample loading platform 12;

[0092] The hot cover device 4 is installed on the frame;

[0093] The reagent tube 5 is installed on the insertion hole and covered by the hot cover device 4. The reagent tube contains a first reagent and a reagent storage tube 54 separated from the first reagent. The reagent storage tube 54 contains a second reagent.

[0094] The position of the reagent tube 5 is adapted to the position of the temperature control device 2 (the adaptation mentioned here means that after the reagent tube 5 is installed, one end of the reagent tube is located in the temperature control device, and the temperature control device can control the temperature in the reagent tube.); the detection position of the fluorescence detection device 3 is directly opposite to the reagent tube 5;

[0095] Detection methods include:

[0096] Step 1: Add the sample to the reagent tube to mix the sample with the first reagent, open the hot cover device 4, put the reagent tube into the insertion hole and cover the hot cover device,

[0097] Step 2: The temperature control device 2 heats and incubates the reaction site of the reagent tube to release the nucleic acid in the sample into the reagent tube.

[0098] Step 3: Add the second reagent in the reagent storage tube to the reagent tube and mix it with the sample.

[0099] Step 4: Use the temperature control device 2 to control the temperature of the reaction part of the reagent tube so that the mixture of the sample and reagent in the reagent tube reaches the first temperature and the nucleic acid is melted at the first temperature;

[0100] Step 5: Use the temperature control device 2 to control the temperature of the reaction part of the reagent tube so that the sample and reagent mixture in the reagent tube reaches the second temperature, and the nucleic acid is replicated at the second temperature;

[0101] Step 6: Use the temperature control device 2 to control the temperature of the reaction part of the reagent tube, so that the temperature of the sample reagent mixture in the reagent tube cycles between the first temperature and the second temperature until the amplification of the nucleic acid is completed;

[0102] Step seven: The fluorescence detection device 3 performs optical detection on the nucleic acid in the reagent tube 5 .

[0103] A display screen 6 may be installed on the housing. The fluorescence detection device 3 feeds back the optical detection result to the control system, and the control system outputs the optical detection result to the display screen 6 .

[0104] The method further includes step A: heating the thermal cover assembly; step A is performed simultaneously with step 6 or before step 6. Heating the thermal cover can prevent uneven heating of the reagent chamber and avoid condensation. It is understood that heating the thermal cover simultaneously with step 6 or before step 6 can achieve this effect, and the best effect is achieved when heating the thermal cover is completed during step 4 or before step 4.

[0105] The working principle of the present invention is:

[0106] Add the sample to the reagent tube to mix the sample with the first reagent, open the hot cover device 4, put the reagent tube into the insertion hole and cover the hot cover device,

[0107] Heat the hot cover device,

[0108] The temperature control device 2 heats and incubates the reaction part of the reagent tube, releasing the nucleic acid in the sample into the reagent tube.

[0109] Add the second reagent in the reagent storage tube to the reagent tube and mix it with the sample.

[0110] The temperature of the reaction part of the reagent tube is controlled by the temperature control device 2 so that the mixture of the sample and reagent in the reagent tube reaches a first temperature and the nucleic acid is melted at the first temperature;

[0111] The temperature of the reaction portion of the reagent tube is controlled by the temperature control device 2 so that the sample and reagent mixture in the reagent tube reaches a second temperature, and the nucleic acid is replicated at the second temperature;

[0112] The temperature of the reaction portion of the reagent tube is controlled by the temperature control device 2, so that the temperature of the sample reagent mixture in the reagent tube cycles between the first temperature and the second temperature until the amplification of the nucleic acid is completed;

[0113] The fluorescence detection device 3 performs optical detection on the nucleic acid in the reagent tube 5;

[0114] The fluorescence detection device 3 feeds back the optical detection result to the control system, and the control system outputs the optical detection result to the display screen.

[0115] The present invention directly adds the sample into a dedicated reagent tube, and places the reagent tube in a specific environment through a hot cover device. The hot cover device and the temperature control device 2 create a reaction environment for the reagent tube, and the second reagent can be added without opening the hot cover, thereby completing the entire polymerase chain reaction process.

[0116] Since the entire polymerase chain reaction process is fully enclosed and pollution-free, does not require a dedicated laboratory environment, and only requires sample addition operations, without the need for professional operators, the present invention can be applied to various scenarios and has a wide range of applications.

[0117] The reaction process of the present invention is also relatively simple, the reaction time is short, the test results can be obtained quickly, and the device of the present invention is compact and can be carried in a bag. These advantages make the present invention suitable for bedside testing and further expand the scope of application of the present invention.

[0118] Since the various parts of the present invention are highly integrated and independent of each other, that is, a reagent tube, a hot cover device, a temperature control device and an optical detection device can be used together on the rack to perform a complete detection process; therefore, the structure of the present invention can be expanded according to the detection requirements and transformed into a multi-channel nucleic acid detection analyzer, such as Figure 1 As shown, two reagent tubes, two thermal cover devices, two temperature control devices, and two optical detection devices are installed in a rack to form a dual-throughput (i.e., simultaneous detection of two samples) nucleic acid detection analyzer. Similarly, utilizing the principles of the present invention, the number of samples that can be simultaneously detected by the nucleic acid detection analyzer can be increased, such as to three, four, or eight channels, allowing the present invention to flexibly adjust the number of detection channels according to different needs.

[0119] Temperature control device

[0120] This section details the specific structure of the temperature control part and how the temperature control device controls the temperature inside the reagent tube.

[0121] like Figure 4 、 Figure 5 As shown, the temperature control device 2 includes a driving component, a guide component, and a first heating component.

[0122] The drive assembly includes a temperature-controlled drive motor 21 mounted on the frame, a bearing seat 22, a lead screw 23 with one end connected to the temperature-controlled drive motor 21 and the other end connected to the bearing seat 22, and a lead screw nut 24 connected to the lead screw 23;

[0123] The guide assembly includes two guide rails 25 mounted on the frame;

[0124] The first heating assembly includes a movable platform 27 mounted on the lead screw nut 24, and a slider 26 is mounted on the movable platform 27 and is in sliding engagement with the guide rail 25;

[0125] There are four heating slots for heating on the mobile platform 27. The four heating slots are arranged parallel to the guide rail 25. Each heating slot is installed with a ceramic heating plate 28 and a heat-conducting copper plate 29 that is tightly against the ceramic heating plate 28. There are two heat-conducting copper plates 29 in each heating slot. There is a gap between the two heat-conducting copper plates 29 for the reagent tube 5 to pass through. The heat-conducting copper plate 29 is in contact with the outer wall of the reagent tube 5.

[0126] The temperatures in the four heating tanks are all different. The temperatures in the four heating tanks include a first target temperature and a second target temperature for polymerase chain reaction, and a first transition temperature and a second transition temperature for transition, with the values of the first transition temperature, the first target temperature, the second target temperature, and the second transition temperature decreasing in sequence.

[0127] The movable platform 27 is made of heat-insulating material. The movable platform 27 is assembled by two interconnected heat-insulating plates. The temperature-controlled drive motor 21 is a stepping motor.

[0128] A trigger member 210 is installed on the mobile platform 27, and an induction switch is installed on the frame, and the positions of the trigger member 210 and the induction switch are adapted; or a trigger member 210 is installed on the frame, and the induction switch is installed on the mobile platform 27, and the positions of the trigger member 210 and the induction switch are adapted (the adaptation mentioned here means that the relative movement of the trigger member 210 and the induction switch can control the induction switch to be triggered).

[0129] A polymerase chain reaction method comprising the following steps:

[0130] Step 1: placing the reagent tube containing the nucleic acid at a first transition temperature and heating it;

[0131] Step 2: placing the reagent tube containing the nucleic acid at a first target temperature to perform a melting reaction;

[0132] Step 3: Place the reagent tube after the melting reaction at the second transition temperature and cool it down;

[0133] Step 4: placing the cooled reagent tube at a second target temperature to replicate the nucleic acid;

[0134] Step 5: Repeat steps 1 to 4;

[0135] Step 6: Repeat step 1 once, or repeat steps 1 to 4 once;

[0136] In the above steps, the values of the first transition temperature, the first target temperature, the second target temperature, and the second transition temperature decrease in sequence.

[0137] In another embodiment, a polymerase chain reaction method comprises the following steps:

[0138] Step 1: Place the reagent tube containing nucleic acid at the first target temperature to perform a melting reaction;

[0139] Step 2: Place the reagent tube after the melting reaction at the second transition temperature and cool it down;

[0140] Step 3: placing the cooled reagent tube at a second target temperature to replicate the nucleic acid;

[0141] Step 4: placing the reagent tube after nucleic acid replication at the first transition temperature and heating it;

[0142] Step 5: Repeat steps 1 to 4;

[0143] Step 6: Repeat steps 1 to 3, or repeat steps 1 to 4.

[0144] In the above steps, the values of the first transition temperature, the first target temperature, the second target temperature, and the second transition temperature decrease in sequence.

[0145] The first transition temperature is at least five degrees Celsius higher than the first target temperature, and the second transition temperature is at least ten degrees Celsius lower than the second target temperature.

[0146] In step 1, the difference between the temperature in the reagent tube after cooling and the second target temperature is at most ten degrees Celsius.

[0147] In step three, the difference between the temperature in the reagent tube after heating and the first target temperature is at most ten degrees Celsius.

[0148] In step 1, the temperature in the reagent tube after cooling is at most five degrees Celsius higher than the second target temperature.

[0149] In step three, the temperature in the reagent tube after heating is at most five degrees Celsius lower than the first target temperature.

[0150] Working principle of temperature control device:

[0151] The ceramic heating plates 28 in different heating tanks are set to different temperatures, and the temperatures are transferred to the heat-conducting copper plate 29 , so that the temperatures of the heat-conducting copper plate 29 in different heating tanks are different.

[0152] After the reagent tube is installed on the sample loading platform, a portion of the reagent tube 5 is located in the gap between the two heat-conducting copper sheets 29. The copper sheets and the reagent tube 5 exchange heat to increase or decrease the temperature inside the reagent tube.

[0153] The temperature-controlled drive motor 21 of the present invention rotates, driving the screw 23 to rotate, and the screw nut 24 drives the movable platform 27 to move on the guide rail 25, so that the reagent tube 5 is placed in different heating tanks. The temperatures in different heating tanks are different, so that the reagent tubes reach different temperatures, realizing polymerase chain reaction.

[0154] The reagent tube 5 is fixed and can be switched between different temperature zones by the reciprocating motion of the mobile platform. This method maintains the reagent tube 5 and the reagents therein in a stable state, which is conducive to the consistency of each reaction. At the same time, compared with heating or cooling at the same position, because the reagent tube 5 directly enters the different temperature zones, the stabilization speed is faster.

[0155] The number of heating tanks is at least three, which is conducive to further improving the temperature change speed. The specific principle is as follows:

[0156] There are two temperatures used for the reaction: the first target temperature for melting and the second target temperature for replication. Therefore, at least two heating tanks are required, one to maintain the first target temperature and the other to maintain the second target temperature. Since the speed of heat conduction is related to the temperature difference, when the temperature difference is relatively small, especially when the temperature is about to reach the target temperature, the speed of heat conduction decreases significantly. This leads to slow temperature switching and prolongs the reaction time. Since a cycle requires at least two temperature switches and the entire reaction process requires dozens of cycles, this problem has a significant impact on the entire reaction process.

[0157] The values of the first target temperature, the second target temperature, and the second transition temperature decrease in sequence. When the temperature of the reagent tube needs to be switched from the first reaction temperature to the second reaction temperature, the reagent tube is first placed in an environment of the second transition temperature to reduce its temperature to the second reaction temperature, and then the reagent tube is placed in an environment of the second reaction temperature to stabilize it at the second reaction temperature. If there are three or more heating tanks, the temperatures of the three heating tanks can be set to the first reaction temperature, the second reaction temperature, and the second transition temperature, respectively, to achieve this process. Due to the large temperature difference between the first target temperature and the second transition temperature, the heat conduction speed is faster, which increases the speed of temperature switching and thus shortens the reaction time.

[0158] Similarly, when the temperature of the reagent tube needs to be switched from the second reaction temperature to the first reaction temperature, the reagent tube is first placed in an environment of the first transition temperature to raise its temperature to the first reaction temperature, and then the reagent tube is placed in an environment of the first reaction temperature to stabilize it at the first reaction temperature.

[0159] In this embodiment, there are four heating tanks, and the temperatures in the four heating tanks are respectively: a first reaction temperature, a second reaction temperature, a first transition temperature, and a second transition temperature. The above temperature changing method can achieve rapid temperature change and multiple cycles.

[0160] Two heat-conducting copper sheets 29 are used to heat or cool down both sides of the reagent tube, which can also increase the speed of temperature change.

[0161] The movable platform 27 is made of heat-insulating material, so that the reagent tube can exchange heat only through the heat-conducting copper sheet, which can increase the temperature change speed and help maintain the temperature in the heating tank stable.

[0162] The thermally conductive copper sheet is attached to the reagent tube so that the heat of the thermally conductive copper sheet is directly transferred to the reagent tube, thus avoiding the problems of slow speed and large heat loss caused by air conduction.

[0163] The temperature-controlled drive motor is a stepping motor, which can control the moving distance of the mobile platform through the temperature-controlled drive motor; the trigger member 210 can accurately feedback the position of the mobile platform, thereby positioning the mobile platform to the initial position.

[0164] Different heating tanks are set up independently, and there is no heat transfer between the heating tanks. The reagent tubes and the heating tanks are also set up independently, which is stable, will not cause pollution, and is simple to maintain, or even maintenance-free.

[0165] The temperature control device of the present invention has a compact structure and a small size.

[0166] The bearing seat 22, temperature-controlled drive motor 21, lead screw 23 and guide rail 25 are all mounted on the lower surface of the sample loading platform 12. This installation method can save space, and the reagent tube, guide rail 25 and lead screw 23 are all positioned with the sample loading platform 12 as the reference, which can improve the relative position accuracy among the three.

[0167] The motion mode of the mobile platform of this embodiment is: the motor drives the lead screw 22 to rotate, and then drives the mobile platform 27 to move linearly through the lead screw nut. Other modes can also be adopted, such as using a lead screw motor or a linear motor to directly drive the mobile platform to slide on the guide rail 25.

[0168] The support frame 13 is made of aluminum, and the center of the support frame 13 is hollow.

[0169] Aluminum products are strong and light, and the hollow center can further reduce its weight.

[0170] Hot cover device

[0171] This section details the specific structure of the thermal cover, the function of the thermal cover, and the role of the thermal cover in instrument automation.

[0172] like Figure 6 、 Figure 7 、 Figure 8 、 Figure 9 As shown, the hot cover device 4 includes a cover shell 41, a second heating assembly installed on the cover shell 41, a guide structure, and a push rod structure.

[0173] The cover shell 41 includes an upper cover shell 411 and a lower cover shell 412 that is matched with the upper cover shell 411. Both the upper cover shell 411 and the lower cover shell 412 are hollow structures. The upper end of the upper cover shell 411 is sealed, and the inner wall of the lower cover shell 412 has a first installation step 413 and a second installation step 414.

[0174] The second heating assembly includes a heat conducting plate 42 and a heating element in contact with the heat conducting plate 42. A first push rod through hole is provided on the heat conducting plate. The lower surface of the heat conducting plate 42 has a downwardly protruding annular boss 421. The shape of the annular boss 421 is adapted to the end of the reagent tube (the adaptation mentioned here means that the annular boss 421 can surround the end of the reagent tube so that the heating space covers the end of the reagent tube). The inner wall of the annular boss 421 and the lower surface of the heat conducting plate 42 form a heating space for heating the end of the reagent tube;

[0175] The second heating assembly includes a heat conducting plate 42 and a heating element in contact with the heat conducting plate 42. The heat conducting plate is provided with a first ejector hole. A fourth groove is provided on the lower surface of the heat conducting plate 42, forming a heating space for heating the end of the reagent tube. The shape of the fourth groove is adapted to the end of the reagent tube. (The "adaptive" mentioned here means that the fourth groove can surround the end of the reagent tube, so that the heating space covers the end of the reagent tube.)

[0176] Three guide holes are provided on the heat conducting plate 42;

[0177] A sensor mounting slot 422 is provided on the heat conducting plate 42, and a temperature sensor is installed in the sensor mounting slot;

[0178] The heating element is a PI heating film, and the heating element is attached to the upper surface of the heat conducting plate 42;

[0179] The guide structure includes a guide plate 43, three guide shafts mounted on the lower surface of the guide plate 43, and a first elastic member 431 sleeved on the guide shafts. The guide plate 43 is provided with a second push rod through hole and a waist-shaped hole 432.

[0180] The waist-shaped hole 432 is used for passing the wires, and both the heating film and the temperature sensor require wire connections.

[0181] The guide shaft is made of stainless steel. In order to ensure smooth movement, the roughness of its surface is extremely low. The end of the guide shaft is a groove, which is convenient for fixing and limiting with a retaining spring.

[0182] The ejector structure includes an ejector mounting frame 441 mounted on the guide plate 43, a linear motor 442 mounted on the ejector mounting frame 441, and an ejector 443 slidably disposed on the ejector mounting frame 441 and cooperatively connected to a slider of the linear motor 442.

[0183] A linear bearing 444 is mounted on the mounting frame of the ejector rod 441 , and the ejector rod 443 is mounted on the linear bearing 444 ;

[0184] A third groove is provided in the middle of the push rod, and the third groove is matched and connected with the slider of the linear motor.

[0185] The end of the ejector pin has an ejector boss. (The ejector boss is used to pierce the sealing membrane at a position corresponding to the position of the reagent storage tube and engages with the first groove 510 at the top of the reagent storage tube to ensure that the reagent storage tube is subjected to a vertical downward force, thereby pushing the reagent storage tube downward.)

[0186] The guide plate 43 of the guide structure is installed on the first step 413, the heat conducting plate 42 of the second heating component is located on the second step 414, the guide hole on the heat conducting plate 42 is connected with the guide shaft, and the first elastic member 431 is located between the heat conducting plate 42 and the guide plate 43, and the end of the top rod 443 passes through the first top rod through hole and the second top rod through hole and is located in the heating space.

[0187] The principle of the hot cover device is:

[0188] The reagent tube is installed on the sample loading table, and the hot cover is placed on the reagent tube so that the reagent tube is covered in the machine and one end of the reagent tube is located in the heating space. The specific process is: when the lower surface of the heat conducting plate 42 contacts the end of the reagent tube 32, the heat conducting plate 42 will be subjected to a reaction force, so that the heat conducting plate 42 moves along the guide shaft toward the guide plate 43, and the first elastic member 431 is compressed until the hot cover reaches the specified position. Since the first elastic member is in a compressed state, the elastic force pushes the heat conducting plate 42 to press the reagent tube, ensuring that the reagent tube 5 is positioned at the specified position so that the reagent tube 5 can be heated, reacted and detected. The hot cover and the sample loading table adopt a commonly used method that can be opened and closed, such as using a snap or magnetic method to ensure that the hot cover is covered on the sample loading table.

[0189] Once the heated cover reaches the designated position, the heating element heats up, applying heat to the heat conducting plate 42, maintaining the desired temperature within the heating chamber. This temperature allows the sample to react, exposing the nucleic acids in the sample. Furthermore, since the entire reagent tube is kept at a relatively high temperature, condensation is effectively avoided. The heating chamber temperature is fed back by a first temperature sensor, typically maintained between 90 and 110 degrees Celsius, effectively preventing condensation.

[0190] When the second reagent needs to be added, the linear motor 442 moves to drive the push rod 443 to slide in the linear bearing 444. The push rod boss will pierce the sealing film at the position corresponding to the position of the reagent storage tube and push the reagent storage tube downward, so that the second reagent moves downward into the reaction liquid; then the push rod 443 returns to its original position.

[0191] After the test is completed, open the hot cover and take out the reagent tube to complete the entire test process.

[0192] The beneficial effects of the hot cover device are:

[0193] Since there is a heat conducting plate 42 for covering the end of the reagent tube, and the heat conducting plate 42 can be heated, it is ensured that the reagent tube 5 can perform amplification reaction and optical detection, and at the same time it will not condense due to uneven heating.

[0194] Because the heat conducting plate 42 can slide and, under the elastic force of the first elastic member, compress the reagent tube, the reagent tube can be compressed, ensuring that the other end of the reagent tube is located within the temperature control device 2 and is in close contact with the fluorescence detection device 3, thereby providing protection for subsequent reactions and detection. At the same time, because the heat conducting plate 42 can slide, it can accommodate reagent tubes of different heights, ensuring that reagent tubes of different heights can all be compressed. This structure greatly reduces the processing precision requirements of the reagent tube and the heating plate.

[0195] Since the hot cover device has a push rod, and the push rod 443 can reciprocate to pierce the sealing film, the reagent storage tube is pushed to move and the second reagent is added. This process does not require manual operation and is completely completed by the instrument. There is no need to open the hot cover, which completely solves the problem of sample contamination during the reaction process. The entire reaction process is carried out in a stable environment, which is beneficial to the stability of the reaction and the accuracy of the test results.

[0196] The nucleic acid detection analyzer originally needs to be provided with a cover for adding reagent tubes or samples into the instrument. The present invention integrates the reagent tube pressing device, heating device and automatic addition device of the second reagent into the cover, which greatly saves space, simplifies the instrument structure, simplifies the operation steps, and thus improves the user experience.

[0197] Since there are many structures inside the thermal cover device, dividing the cover shell 41 into two parts is beneficial for the installation of the internal structures of the thermal cover device.

[0198] The first installation step 413 and the second installation step 414 are relatively easy to process, and the remaining structures on the first installation step 413 and the second installation step 414 are relatively easy to process.

[0199] The heating space is generated by using the annular boss 421 or the fourth groove. These two methods are relatively simple to process and do not require the installation of other parts. In addition, these two methods are conducive to the heating element to conduct heat into the heating space.

[0200] Using PI heating film for heating can avoid processing a structure dedicated to installing heating elements on the heat conducting plate.

[0201] The guide plate 43 is used to install the ejector structure and guide the second heating assembly.

[0202] Since the heat conducting plate 42 is guided by the guide shaft, the heat conducting plate 42 is limited in the axial direction of the guide shaft by the second step, which greatly reduces the processing accuracy requirements and the installation difficulty. The specific reason is: if the limiting structure of the heat conducting plate 42 in the axial direction of the guide shaft is set on the guide shaft, it is necessary to ensure the matching accuracy of the guide shaft and the guide hole, as well as the position accuracy and installation accuracy of the limiting structure. The processing accuracy requirements will be very high, and since the internal space of the thermal cover device is relatively small, the installation is difficult; similarly, if the structure for guidance is set on the cover shell, the processing accuracy requirements of the cover shell will be very high. Considering that the structure of the cover shell is relatively thin, the processing difficulty will be even greater; in this solution, only the matching accuracy of the guide shaft and the guide hole needs to be considered, and only the position accuracy of the limiting structure on the cover shell needs to be ensured, and the processing difficulty is very low.

[0203] A linear motor 442 is used to drive the ejector pin, and the slider of the linear motor engages with the third groove of the ejector pin 443. This transmission method is simple and low-cost. Other commonly used transmission structures can also be used to replace the above transmission mechanism, such as a rotary motor in conjunction with a gear rack, a worm gear, or a lead screw slide; or an air pump or hydraulic pump can be used to drive the ejector pin.

[0204] Reagent tube

[0205] This section describes in detail the specific structure of the reagent tube, how to automatically mix the first reagent and the second reagent in the reagent tube, and other functions of the reagent tube.

[0206] like Figure 10 、 Figure 11 As shown, a reagent tube includes a tube cover 51 and a reaction tube 52. The tube cover 51 is provided with a sample addition hole and a reagent addition hole; a reagent storage tube 54 is installed in the reagent addition hole, and the reagent storage tube is filled with a second reagent. The tube cover 51 has a sealing film for sealing the sample addition hole and the reagent addition hole; the reaction tube 52 has a reaction chamber 53; the tube cover 51 and the reaction tube 52 are connected to each other so that the reaction chamber is in a sealed state.

[0207] The cross section of the tube cover 51 is circular or square. The circular or square shape is convenient for gripping. A reagent tube positioning groove is provided on the sample loading platform, and a first protrusion 56 is provided on the outer wall of the tube cover, which is adapted to the reagent tube positioning groove.

[0208] A rubber sealing plug 55 is fixed in the sample adding hole. A second protrusion is provided on the inner wall of the sample adding hole, and a second groove is provided on the rubber sealing plug 55 to match the second protrusion.

[0209] The reagent storage tube 54 is movably installed in the reagent adding hole of the tube cover 51.

[0210] The reagent storage tube is tubular, with one end of the reagent storage tube being closed and the other end being open.

[0211] The reagent storage tube 54 has an inclined edge 59 at its open end and a first groove 510 at its closed end.

[0212] The reagent storage tube 54 further includes a gravity ball 57 . The second reagent is a freeze-dried ball 58 . The gravity ball 57 is placed above the freeze-dried ball 58 .

[0213] The gravity ball is a solid with a smooth surface and is made of polymer materials.

[0214] The reagent storage tube 54 further includes a second elastic member. The second reagent is a freeze-dried ball 58 . The lower end of the second elastic member abuts against the freeze-dried ball, and the upper end of the second elastic member abuts against the reagent storage tube.

[0215] The second elastic member is a plastic spring.

[0216] The plastic spring is pre-compressed in the reagent storage tube to reduce the vibration of the freeze-dried balls during transportation. When the sealing film on the reaction tube is punctured, it can give the freeze-dried balls an ejection force, allowing them to fall into the solution more smoothly.

[0217] Both end openings of the sample adding hole and both end openings of the reagent adding hole are provided with sealing films, and the sealing films can be puncture-prone films.

[0218] The sample wells and reagent wells are both circular through holes.

[0219] The reaction tube contains a first reagent.

[0220] The lower half of the reaction tube 52 is flat and has a transparent detection window.

[0221] The detection window is located at the bottom of the reaction tube 52 .

[0222] The upper half of the reaction tube 52 has a circular or rectangular cross-section. The upper half of the reaction tube 52 is connected to the tube cover 51. A sealing film is provided at the opening of the reaction tube 52 to seal one end of the sample addition hole and the reagent addition hole. A sealing film is provided on the upper surface of the tube cover 51 to seal the other end of the sample addition hole and the reagent addition hole. The circular or rectangular shape is easy to process and convenient to take and put. In addition, the circular tube wall can ensure uniform thickness, which is conducive to ensuring that the temperature inside the reaction tube is stable at the target value through heating. This structure has a wider upper portion, which can ensure that the reagent and sample are fully in contact and react, and ensure the dissolution of the freeze-dried ball. The narrower lower structure ensures the heat conduction speed and PCR amplification speed.

[0223] Working principle of the reagent tube: When adding samples, the syringe pierces the seal on the tube cover 51, the rubber sealing plug and the sealing film on the reaction tube 52, enters the first reagent for adding samples, and when the syringe is removed, the rubber sealing plug closes itself due to elasticity.

[0224] Because the reaction tube contains the first reagent, the sample can react with the first reagent first. Since the tube cap contains the second reagent, the second reagent can be added to the reaction tube at the appropriate time to carry out the next reaction on the sample. After the reaction is completed, the detection device detects it through the detection window at the bottom of the reaction tube.

[0225] The working principle of adding the second reagent to the reaction tube is: push the reagent storage tube, the inclined edge of the reagent storage tube will pierce the sealing film on the reaction tube, so that the second reagent enters the reagent reaction tube and mixes with the first reagent and the sample.

[0226] A push rod can be provided on the instrument, and the reagent tube is installed in the reaction chamber. The push rod can be moved to push the reagent storage tube.

[0227] The second reagent in this embodiment is stored in the form of freeze-dried pellets. The advantages of freeze-dried pellets are:

[0228] When the oblique blade of the reagent storage tube pierces the sealing film, both the freeze-dried ball and the gravity ball will fall into the reaction tube, and the gravity of the gravity ball will ensure that the freeze-dried ball can fall and completely penetrate into the first reagent.

[0229] After the freeze-dried ball is completely dissolved, the instrument will heat the thin wall at the bottom of the reaction tube to start the PCR reaction, and finally perform detection through the detection window at the bottom of the reaction tube.

[0230] The beneficial effects of the reagent tube are:

[0231] Since the reagent tube of the present invention contains both the first reagent and the second reagent, and can react with the sample separately, nucleic acid extraction and PCR amplification reactions can both be performed in the reagent tube, avoiding the transfer of nucleic acids and significantly reducing the cost and complexity of the instrument;

[0232] Since the second reagent can be added without opening the reagent tube, the operation steps are reduced and the risk of sample contamination is also reduced;

[0233] Since the sample addition hole of the reagent tube is provided with a rubber sealing plug, it is convenient for a common syringe to add samples, and after adding the samples, the elasticity of the rubber plug automatically seals the sample, thereby improving the sealing performance of the reaction tube.

[0234] The sealing membrane of the sample well and reagent well can ensure that the inside of the reagent tube is in a sealed state to avoid contamination and improve the accuracy of detection;

[0235] The second protrusion cooperates with the second groove of the rubber sealing plug 55 to ensure that the rubber sealing plug 55 does not slide in the sample loading hole, thereby ensuring that the puncture of the sample loading needle can proceed smoothly and avoiding the situation where the rubber sealing plug 55 moves and damages the sealing membrane.

[0236] The first protrusion on the outer wall of the tube cover is used to cooperate with the instrument to achieve positioning, facilitate installation, and ensure that the reagent tube can be installed in the designated position for reaction and detection.

[0237] The reagent storage tube is used to store and release the second reagent. To release the second reagent, the tip of the beveled blade punctures a point on the sealing membrane. As the reagent storage tube continues to move, the edge of the beveled blade tears the sealing membrane, allowing the reagent storage tube to pass through the sealing membrane and allowing the second reagent to enter the reaction tube. The sealing membrane is only punctured at the point where the beveled blade contacts the tip of the blade. The membrane distal to the tip remains intact and remains connected to the original membrane, ensuring that the sealing membrane does not escape into the reagent.

[0238] The use of freeze-dried balls in conjunction with reagent storage tubes eliminates the need to worry about the easy flow and leakage of liquid reagents, and eliminates the need for ineffective frozen storage. This makes it easier to load, seal, store, and transport the second reagent, improving the convenience of the reagent tubes and instruments.

[0239] The gravity ball, placed above the lyophilized balls, fills the gaps in the reagent storage tube, preventing the balls from breaking due to vibration in the tube during transport. The gravity ball uses gravity to press down on the lyophilized balls, allowing them to fall smoothly from the reagent storage tube. Once they fall, they hold the balls down, increasing the contact area between the lyophilized balls and the first reagent, ensuring full dissolution of the reagent.

[0240] The lower part of the reaction tube is designed to be flat so that the heating module can fit perfectly, and the thin-wall structure here increases the heating efficiency, ultimately significantly shortening the detection time.

[0241] The PCR detection reagent tube has a simple overall structure and a small size.

Claims

1. A nucleic acid detection analyzer, comprising a frame, characterized in that: It also includes a temperature control device, a fluorescence detection device, a hot cover device and reagent tubes installed on the rack; The frame includes a bottom plate and a sample loading platform mounted on the bottom plate, wherein the sample loading platform is provided with an insertion hole; The reagent tube includes a tube cover and a reaction tube, wherein the tube cover is provided with a reagent adding hole; a reagent storage tube is installed in the reagent adding hole, and the reagent storage tube is movably installed in the reagent adding hole of the tube cover; the reagent storage tube is filled with a second reagent, and the reaction tube has a reaction cavity; the reagent tube is installed on the insertion hole; The temperature control device and the fluorescence detection device are installed between the bottom plate and the sample loading platform. The position of the temperature control device is adapted to the position of the reagent tube and is used to adjust the temperature in the reaction chamber of the reaction tube; The detection position of the fluorescence detection device is directly opposite to the reagent tube; The temperature control device includes a movable platform for heating the lower end of the reagent tube, the movable platform is located between the sample loading platform and the fluorescence detection device, and the temperature control device includes a drive component, a guide component, and a first heating component; the first heating component includes a movable platform, and the movable platform has a plurality of heating slots for heating. The end of the reagent tube is located in a heating slot. The movable platform moves along the guide component under the drive of the drive component, so that the end of the reagent tube is located in different heating slots; The hot cover device is installed on the frame and is used to cover the reagent tube; the hot cover device also includes a push rod structure for pushing the reagent storage tube to move and add the second reagent in the reagent storage tube into the reaction tube; The thermal cover device includes a cover shell and a second heating assembly mounted on the cover shell; the second heating assembly includes a heat conducting plate and a heating element in contact with the heat conducting plate; the lower surface of the heat conducting plate has a downwardly protruding annular boss or the lower surface of the heat conducting plate is provided with a fourth groove; the shape of the annular boss is adapted to the end of the reagent tube, and the inner wall of the annular boss and the lower surface of the heat conducting plate enclose a heating space for heating the end of the reagent tube; the shape of the fourth groove is adapted to the end of the reagent tube and forms a heating space for heating the end of the reagent tube; A first push rod through hole is provided on the heat conducting plate; The push rod structure includes a push rod mounting frame installed on the cover shell, a push rod driving member installed on the push rod mounting frame, a push rod slidably arranged on the push rod mounting frame and cooperatively connected to the push rod driving member, and the driving member can drive the end of the push rod into the heating space.

2. The nucleic acid detection analyzer according to claim 1, wherein: One end of the reagent tube covered by the hot cover is located above the sample loading platform, and the other end is located below the sample loading platform. The fluorescence detection device is located below the reagent tube.

3. The nucleic acid detection analyzer according to claim 1, wherein: The number of the temperature control device, the fluorescence detection device, the hot cover device and the reagent tubes is at least two, and the temperature control device, the fluorescence detection device, the hot cover device and the reagent tubes are matched one by one.

4. The nucleic acid detection analyzer according to claim 1, wherein: The frame further comprises a support frame and a shell mounted on the bottom plate, the sample loading platform is mounted on the support frame, and the hot cover device is mounted on the sample loading platform.

5. The nucleic acid detection analyzer according to claim 1, 2, 3 or 4, characterized in that: The mobile platform is provided with more than three heating tanks for heating.

6. The nucleic acid detection analyzer according to claim 5, characterized in that: There are four heating tanks, which are arranged in parallel with the guide rails. A ceramic heating plate and a heat-conducting copper plate tightly abutting the ceramic heating plate are installed in each heating tank. There are two heat-conducting copper plates in each heating tank, and a gap is provided between the two heat-conducting copper plates for the reagent tube to pass through. The heat-conducting copper plates are in contact with the outer wall of the reagent tube. The temperatures in the four heating tanks include a first target temperature and a second target temperature for polymerase chain reaction, and a first transition temperature and a second transition temperature for transition, and the values of the first transition temperature, the first target temperature, the second target temperature, and the second transition temperature decrease in sequence. The driving assembly includes a temperature-controlled driving motor mounted on a frame, a bearing seat, a screw at one end being connected to the temperature-controlled driving motor and the other end being connected to the bearing seat, and a screw nut being connected to the screw. The guide assembly includes two guide rails mounted on a frame. The movable platform is mounted on the screw nut, and the movable platform is also equipped with a slider slidably connected to the guide rail. The mobile platform is made of heat-insulating material; the mobile platform is composed of two interconnected heat-insulating plates; the temperature-controlled drive motor is a stepping motor; a trigger is installed on the mobile platform, an induction switch is installed on the frame, and the positions of the trigger and the induction switch are adapted to each other; or a trigger is installed on the frame, an induction switch is installed on the mobile platform, and the positions of the trigger and the induction switch are adapted to each other.

7. The nucleic acid detection analyzer according to claim 1, 2, 3, 4, or 6, characterized in that: The heat cover device also includes a guide structure installed on the cover shell, the guide structure includes a guide plate, a guide shaft installed on the lower surface of the guide plate, and a first elastic member, and the first elastic member is located between the heat conducting plate and the guide plate; a guide hole is opened on the heat conducting plate, and the guide hole is cooperatively connected with the guide shaft.

8. The nucleic acid detection analyzer according to claim 7, wherein: The guide plate is provided with a second push rod through hole and a waist-shaped hole.

9. The nucleic acid detection analyzer according to claim 7, wherein: The cover shell includes an upper cover shell and a lower cover shell connected to the upper cover shell. The upper cover shell and the lower cover shell are both hollow structures. The upper end of the upper cover shell is sealed, and the inner wall of the lower cover shell has a first mounting step and a second mounting step; the guide plate is installed on the first step, and the heat conducting plate is located on the second step.

10. The nucleic acid detection analyzer according to claim 1 or 2 or 3 or 4 or 6 or 8 or 9, characterized in that: The tube cover is provided with a sample addition hole and a sealing film for sealing the sample addition hole and the reagent addition hole; the tube cover and the reaction tube are cooperatively connected to keep the reaction chamber in a sealed state.

11. The nucleic acid detection analyzer according to claim 10, wherein: The reagent storage tube is tubular, one end of the reagent storage tube is closed and the other end is open; the open end of the reagent storage tube It has an oblique edge and a first groove is provided at the closed end.

12. The nucleic acid detection analyzer according to claim 10, wherein: The reagent storage tube is further provided with a gravity ball, the second reagent is a freeze-dried ball, and the gravity ball is placed above the freeze-dried ball.

13. The nucleic acid detection analyzer according to claim 1 or 2 or 3 or 4 or 6 or 8 or 9 or 11 or 12, characterized in that: The reaction tube contains a first reagent, and the lower half of the reaction tube has a transparent detection window; the detection window is located at the bottom of the reaction tube.

14. The nucleic acid detection analyzer according to claim 10, wherein: Both the openings at both ends of the sample addition hole and the openings at both ends of the reagent addition hole are provided with sealing films, and both the sample addition hole and the reagent hole are circular through holes; or / and, A reagent tube positioning groove is provided on the sample loading platform, the cross-section of the tube cover is circular or square, and a first protrusion is provided on the outer wall of the tube cover, which is adapted to the reagent tube positioning groove; a rubber sealing plug is fixed in the sample loading hole; and the lower half of the reaction tube is flat.

15. A detection method for a nucleic acid detection analyzer, characterized in that: Detection is performed using the nucleic acid detection analyzer according to any one of claims 1 to 14; The detection method comprises: Step 1: Add the sample to the reagent tube to mix the sample with the first reagent, open the hot cover device, put the reagent tube into the insertion hole and cover the hot cover device, Step 2: The mobile platform of the temperature control device heats and incubates the reaction site of the reagent tube, releasing the nucleic acid in the sample into the reagent tube. Step 3: Add the second reagent in the reagent storage tube to the reagent tube and mix it with the sample. Step 4: Moving the movable platform of the temperature control device to control the temperature of the reaction site of the reagent tube so that the sample and reagent mixture in the reagent tube reaches a first temperature and the nucleic acid is melted at the first temperature; Step 5: Moving the movable platform of the temperature control device to control the temperature of the reaction portion of the reagent tube so that the sample-reagent mixture in the reagent tube reaches a second temperature, and the nucleic acid replicates at the second temperature; Step 6: Move the mobile platform of the temperature control device to control the temperature of the reaction site of the reagent tube, so that the temperature of the sample reagent mixture in the reagent tube cycles between the first temperature and the second temperature until the nucleic acid amplification is completed; Step 7: The fluorescence detection device performs optical detection on the nucleic acid in the reagent tube.

16. The detection method of the nucleic acid detection analyzer according to claim 15, characterized in that: The method further comprises step A: heating the hot cover device; step A is performed simultaneously with step 6 or before step 6.

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