Automatic plasma sample detection device and method based on integrated micro-fluidic chip

Abstract

The invention discloses an automatic plasma sample detection device and method based on an integrated micro-fluidic chip. The device comprises a chip action mechanism, a magnetic bar operation mechanism, a pipette operation mechanism and a fluorescence detection mechanism which are arranged in a device base, wherein the magnetic bar operating mechanism, the pipette operating mechanism and the fluorescence detection mechanism are respectively opposite to the top and the side surface of the chip actuating mechanism. Through cooperative control of the chip, the magnetic bar / magnetic bar sleeve movement mechanism and the pipetting mechanism, the device can be greatly simplified, the operation complexity is reduced, the detection time is shortened, and super-multiple detection for detecting more than 30 biological targets at one time is realized.

Description

Technical Field

[0001] The present invention relates to a technology in the field of biological detection, specifically to an automated detection device and method for plasma samples based on an integrated microfluidic chip. Background Art

[0002] The current nucleic acid detection technology has a long process, complex operation, large related equipment and high cost. Sample processing, nucleic acid extraction, purification, amplification and detection equipment are mostly independent devices. Samples need to be transferred between multiple devices, which can easily cause contamination. In addition, the number of detection targets is small, which limits its application scenarios. Summary of the invention

[0003] In view of the above-mentioned deficiencies in the prior art, the present invention proposes an automated detection device and method for plasma samples based on an integrated microfluidic chip. Through the coordinated control of the chip, the magnetic rod / magnetic rod sleeve motion mechanism, and the pipetting mechanism, the equipment can be greatly simplified, the operation complexity can be reduced, the detection time can be shortened, and ultra-multiple detection of more than 30 biological targets can be achieved at one time.

[0004] The present invention is achieved through the following technical solutions:

[0005] The present invention relates to an automated plasma sample detection device, comprising: a chip action mechanism, a magnetic rod operation mechanism, a pipette operation mechanism and a fluorescence detection mechanism arranged in a device base, wherein: the magnetic rod operation mechanism, the pipette operation mechanism and the fluorescence detection mechanism are respectively opposite to the top and the side of the chip action mechanism.

[0006] The chip action mechanism includes: a motor bracket, a chip rotating motor, an integrated microfluidic chip, and a heating film bracket, a heating film and a heat-conducting aluminum ring arranged outside the motor bracket in sequence from bottom to top, wherein: the top of the heat-conducting aluminum ring is in contact with the outer bottom of the integrated microfluidic chip.

[0007] The integrated microfluidic chip comprises: a motor matching ring located in the center, a pipette tip pre-storage chamber arranged on the bottom surface of the integrated microfluidic chip and located on its periphery, a magnetic rod sleeve pre-storage chamber, a magnetic bead liquid pre-storage chamber, a lysis solution pre-storage chamber, two washing solution pre-storage chambers, an eluent pre-storage chamber, a diluent pre-storage chamber, and a sample addition area, a siphon valve, a plurality of parallel test units and a waste liquid chamber arranged on the top surface of the integrated microfluidic chip, wherein: each test unit comprises: an array liquid separation groove, a capillary valve and a reaction chamber, wherein: a motor is connected to the motor matching ring to drive the chip to rotate, so that each chamber can be aligned with the magnetic rod and the pipette, that is, coaxial in the vertical direction.

[0008] The fluorescence detection mechanism includes: an optical path fixture, a collimating lens, a laser transmitter, an optical fiber connected to the collimating lens, a fluorescence receiver, and a circuit board, wherein: after the incident laser triggers the fluorescence reaction of the reagent in the chip detection area, it is received by the collimating lens and transmitted to the fluorescence receiver through the optical fiber. In the fluorescence receiver, the light path passes through the filter and is received by the fluorescence sensor for subsequent analysis.

[0009] The fluorescence detection mechanism further comprises a thin film temperature sensor connected to the circuit board, and the thin film temperature sensor is arranged on the heating film of the chip action mechanism. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] Figure 1 is a schematic diagram of the present invention;

[0011] Figure 2 It is a top view of the integrated chip;

[0012] Figure 3 It is a bottom view of the integrated chip;

[0013] Figure 4 It is a schematic diagram of the magnetic rod / magnetic rod sleeve motion mechanism of the present invention;

[0014] Figure 5 It is a schematic diagram of the fluorescence detection mechanism of the present invention;

[0015] In the figure: device base 1, chip 2, heating film 3, heating film bracket 401, thermal conductive aluminum ring 402, motor bracket 5, chip rotating motor 6, support frame moving motor 7, support frame moving motor slider 8, support frame moving motor screw 9, motor support frame 10, magnetic rod 11, magnetic rod sleeve 12, magnetic rod moving motor 13, pipette moving motor 14, pipette moving motor slider 15, pipette moving motor screw 16, pipette tip 17, pipette 18, pipette moving motor pad 19, optical path fixing part 20, collimating lens 21, optical fiber 22, laser transmitter 23, fluorescence receiver 24, circuit board 25, fixing screw 26, thin film temperature sensor 27. DETAILED DESCRIPTION

[0016] like Figure 1 As shown, a plasma sample automated detection device involved in this embodiment includes: a chip action mechanism, a magnetic rod operation mechanism, a pipette operation mechanism and a fluorescence detection mechanism arranged in the device base 1, wherein: the magnetic rod operation mechanism, the pipette operation mechanism and the fluorescence detection mechanism are respectively opposite to the top and the side of the chip action mechanism.

[0017] The chip action mechanism includes: a motor bracket 5, a chip rotating motor 6, an integrated microfluidic chip 2, and a heating film bracket 401, a heating film 3 and a thermal conductive aluminum ring 402 arranged on the outside of the motor bracket 5 in sequence from bottom to top, wherein: the top of the thermal conductive aluminum ring 402 is in contact with the outer bottom of the integrated microfluidic chip 2.

[0018] like Figure 2 and Figure 3 As shown, the integrated microfluidic chip 2 comprises: a motor matching ring 214 located in the center, a pipette tip pre-storage chamber 201 arranged on the bottom surface of the integrated microfluidic chip 2 and located on its periphery, a magnetic rod sleeve pre-storage chamber 202, a magnetic bead liquid pre-storage chamber 203, a lysis solution pre-storage chamber 204, two washing solution pre-storage chambers 205 and 206, an eluent pre-storage chamber 207, a diluent pre-storage chamber 208, and a sample addition area 209 arranged on the top surface of the integrated microfluidic chip 2, a siphon valve 210, a plurality of parallel test units and a waste liquid chamber 213, wherein: each test unit comprises: an array liquid separation groove 211, a capillary valve 215 and a reaction chamber 212, wherein: a motor 6 is connected to the motor matching ring 214 to drive the chip to rotate so that each chamber can be aligned with the magnetic rod 11 and the pipette 18, that is, coaxial in the vertical direction.

[0019] like Figure 1 and Figure 4 As shown, the magnetic rod operating mechanism includes: a moving motor 7, a moving motor slider 8, a moving motor lead screw 9, and a motor support frame 10, a magnetic rod 11, a magnetic rod sleeve 12, and a magnetic rod moving motor 13 arranged on the moving motor slider 8 in sequence, wherein: the slider 8 is fixedly connected to the support frame 10, and the motor 19 is fixedly connected to the support frame 10. The lead screw 9 rotates to drive the slider 8 to move vertically, and the support frame 10 moves vertically. The inner side of the magnetic rod sleeve 12 is clamped by the magnetic rod sleeve fixing ring 1001 to achieve the installation of the magnetic rod sleeve. The rotation of the lead screw 9 can control the magnetic rod sleeve 11 to enter each chamber to absorb and transfer the magnetic beads; the lead screw 9 repeatedly rotates clockwise and counterclockwise to drive the magnetic rod sleeve 11 to move back and forth in the vertical direction to achieve the beating and mixing of the liquid. The bottom of the telescopic rod of the motor 19 is fixedly connected to the magnetic rod 11. The motor 19 drives the magnetic rod 11 to move vertically. Moving upward to leave the magnetic rod sleeve 12 is demagnetization, and moving downward to push the magnetic rod sleeve 12 out of the fixing ring 1001 is unloading of the magnetic rod sleeve.

[0020] The pipette operating mechanism includes: a pipette moving motor pad 19 fixedly arranged on the device base 1, and a pipette moving motor 14, a pipette moving motor screw 16, a pipette moving motor slider 15, a pipette 18 and a pipette head 17 arranged thereon in sequence.

[0021] like Figure 1 and Figure 5As shown, the fluorescence detection mechanism includes: an optical path fixture 20, a collimating lens 21, a laser transmitter 23, an optical fiber 22 connected to the collimating lens 21, a fluorescence receiver 24, and a circuit board 25, which are arranged in sequence, wherein: after the incident laser triggers the fluorescence reaction of the reagent in the chip detection area 212, it is received by the collimating lens 21 and transmitted to the fluorescence receiver 24 through the optical fiber 22. In the fluorescence receiver 24, the light path passes through the filter and is received by the fluorescence sensor for subsequent analysis.

[0022] The fluorescence detection mechanism further includes a thin film temperature sensor 27 connected to the circuit board 25, and the thin film temperature sensor 27 is arranged on the heating film 3 of the chip action mechanism.

[0023] The present embodiment relates to an automated detection method based on the above system, which includes adding a plasma sample and proteinase K to a chip lysis chamber, installing the chip to a predetermined position of the device, closing the device door, and clicking a start detection button on the touch screen; installing a magnetic rod sleeve according to a preset program, and rotating the chip with a motor so that the magnetic bead pre-storage chamber is aligned with the magnetic rod sleeve, and a magnetic rod motion mechanism drives the magnetic rod / magnetic rod sleeve to enter the magnetic bead pre-storage chamber to absorb the magnetic beads, and after transferring them to the lysis chamber, the magnetic rod is separated from the magnetic rod sleeve, and the plasma sample is lysed under the auxiliary beating action of the magnetic rod sleeve, and the sample nucleic acid is combined with the magnetic beads; the magnetic rod enters the magnetic rod sleeve again, adsorbs the magnetic beads, and is lifted above the chip, and the motor drives the chip to rotate so that the washing chamber is aligned with the magnetic rod sleeve, and the magnetic rod adsorbs the magnetic beads and enters the washing chamber. chamber, complete 2 washings in sequence, after drying the magnetic beads, transfer the magnetic beads to the elution chamber, the elution solution elutes the nucleic acid from the magnetic beads, and the magnetic beads are taken away from the elution chamber by a magnetic rod to obtain the nucleic acid elution solution, and the diluent is transferred to the nucleic acid elution chamber by an automatic pipetting device, and after being fully mixed with the nucleic acid elution solution, the mixed solution is transferred to the mixing chamber of the outer ring of the chip, and then the chip rotates under the control of the motor, and the centrifugal force is used to drive the mixed solution to the liquid separation and metering chamber to complete the equal division and metering of the liquid, and the chip is then rotated at a high speed to break the capillary valve so that the liquid in the metering chamber enters each reaction chamber, and under the action of the temperature control device, the nucleic acid is isothermally amplified in each reaction chamber, and after the isothermal amplification is completed, the amplification product is detected by a fluorescence detection device to obtain the detection result.

[0024] The sample pre-treatment specifically includes:

[0025] Step 101: To avoid interference, the sliders 8 and 15 are moved up to the standby position to leave space.

[0026] Step 102 : After manually adding a plasma sample into the chamber 204 , the matching ring 214 of the chip 2 is installed on the motor 6 .

[0027] Step 103, the chip 2 moves clockwise, and the chamber 202 reaches the bottom of the magnetic bar 11; the support frame 10 moves downward, and after the magnetic bar sleeve 12 is installed, the support frame 10 moves upward to the standby position.

[0028] Step 104, the chip 2 rotates clockwise, and the chamber 203 reaches the bottom of the magnetic rod 11; the support frame 10 moves downward, and after the magnetic rod 11 absorbs the magnetic beads, the support frame 10 moves upward to the standby position.

[0029] Step 105, the chip 2 rotates clockwise, and the chamber 204 reaches the bottom of the magnetic rod 11. At this time, the chamber 204 contains the lysed sample; the support frame 10 moves downward, and the magnetic rod 11 moves upward for demagnetization; then the magnetic rod sleeve 12 starts to oscillate up and down, and the motor 6 drives the chip 2 to rotate clockwise and reversely in a small amplitude, simulating beating and mixing, and continues for five times.

[0030] Step 106, the magnetic rod 11 moves downward, returns to the inside of the magnetic rod sleeve 12, and re-absorbs the magnetic beads.

[0031] Step 107, the support frame 10 moves up and stands by.

[0032] Step 108, the chip 2 moves clockwise, and the first washing chamber 205 is in place; the magnetic rod sleeve 12 moves downward, moves upward and returns after washing.

[0033] Step 109, the chip 2 moves clockwise, and the second washing chamber 206 is in place; the magnetic rod sleeve 12 moves downward, moves upward and returns after washing.

[0034] Step 110, the chip 2 moves clockwise, and the elution chamber 207 is in place; the magnetic rod cover 12 moves downward, and the bottom enters the elution solution and is immersed; after the elution operation is completed, the magnetic rod cover 12 moves up and returns to the standby position.

[0035] Step 111, the chip moves clockwise, and the chamber 202 for placing the magnetic rod sleeve is in place; the magnetic rod sleeve 12 moves down, and the magnetic rod 11 pushes out of the magnetic rod sleeve 12 for unloading, and then the support frame 10 moves up and returns to the standby position.

[0036] The pipetting amplification specifically comprises:

[0037] Step 201, initializing the pipette.

[0038] Step 202, the chip 2 rotates clockwise, and the chamber 201 moves to the position directly below the pipette 18; a pipette tip is removed from the pipette 18, and a tip detection is performed.

[0039] Step 203: The pipette gun 18 moves upward to a standby position.

[0040] Step 204 , the chip 2 rotates clockwise, and the chamber 208 moves to the position directly below the pipette gun 18 ; the pipette gun 18 moves downward, absorbs the diluent in the chamber 208 , and then moves upward to the standby position.

[0041] Step 205 , the chip 2 rotates clockwise, and the chamber 207 moves to the position directly below the pipette gun 18 ; the pipette gun 18 moves downward to discharge the liquid, and then moves upward to the standby position.

[0042] Step 206 : The chip 2 oscillates back and forth, and the sample and the diluent in the chamber 207 are mixed.

[0043] Step 207 , the pipette 18 moves down and aspirates the liquid, and then moves up to the standby position.

[0044] Step 208 , the chip 2 rotates clockwise, and the chamber 209 moves to the position directly below the pipette 18 ; the pipette 18 moves downward and adds the target sample from the sample addition hole into the chamber 209 , and then moves upward to the standby position.

[0045] Step 209: The chip 2 rotates clockwise, and the chamber 201 moves to the position directly below the pipette 18; the pipette performs a tip withdrawal operation.

[0046] The liquid separation detection specifically includes:

[0047] Step 301, the heating film 3 works, the heat-conducting aluminum ring 402 heats up, and temperature feedback is performed through the temperature sensor 27. The heating film 3 starts and stops to control the heat-conducting aluminum ring 402 to be in a constant temperature state, ensuring that the liquid in the outer ring chamber of the chip 2 is in a constant temperature environment of 37±0.5°C.

[0048] Step 302 , the motor 6 rotates slowly back and forth to vibrate and mix the liquid in the chamber 209 .

[0049] Step 303 , reverse the valve and break it, and the liquid enters the siphon valve 210 ; then slowly accelerate to the liquid separation speed, and the liquid evenly enters the array liquid separation tank 211 .

[0050] Step 304 , continue to increase the rotation speed, and the liquid enters the array fluorescence detection area 212 through the capillary valve 215 for fluorescence amplification.

[0051] Step 305, rotate the chip 2 in sequence to detect the fluorescence reaction of each chamber in the array fluorescence detection area 212. The fluorescence is received by the collimating lens and transmitted through the optical fiber. After passing through the filter in the fluorescence receiver 24, the fluorescence sensor collects the signal and the signal is returned to the circuit board to determine the positive and negative.

[0052] The specific speed test results are shown in the following table.

[0053] Compared with the prior art, the integrated microfluidic chip involved in the present invention has an inner chamber center and an outer sample loading hole center located on a concentric circle, and the number of chip reaction chambers exceeds 30. Through the coordinated control of the chip and the magnetic rod / magnetic rod sleeve motion mechanism and the pipetting mechanism, the equipment can be greatly simplified, the operation complexity can be reduced, the detection time can be shortened, and ultra-multiple detection of more than 30 biological targets can be achieved at one time.

[0054] The above-mentioned specific implementation can be partially adjusted in different ways by those skilled in the art without departing from the principle and purpose of the present invention. The protection scope of the present invention shall be based on the claims and shall not be limited by the above-mentioned specific implementation. Each implementation scheme within its scope shall be subject to the constraints of the present invention.

Claims

1. A plasma sample automated detection device, characterized in that: include: A chip action mechanism, a magnetic rod operation mechanism, a pipette operation mechanism and a fluorescence detection mechanism are arranged in the base of the device, wherein: the magnetic rod operation mechanism, the pipette operation mechanism and the fluorescence detection mechanism are respectively facing the top and the side of the chip action mechanism; The integrated microfluidic chip comprises: a motor matching ring located in the center, a pipette tip pre-storage chamber arranged on the bottom surface of the integrated microfluidic chip and located on its periphery, a magnetic rod sleeve pre-storage chamber, a magnetic bead liquid pre-storage chamber, a lysis solution pre-storage chamber, two washing solution pre-storage chambers, an eluent pre-storage chamber, a diluent pre-storage chamber, and a sample addition area, a siphon valve, a plurality of parallel test units and a waste liquid chamber arranged on the top surface of the integrated microfluidic chip, wherein: each test unit comprises: an array liquid separation groove, a capillary valve and a reaction chamber, wherein: a motor is connected to the motor matching ring to drive the chip to rotate, so that each chamber can be aligned with the magnetic rod and the pipette, that is, coaxial in the vertical direction.

2. The plasma sample automated detection device according to claim 1, characterized in that: The chip action mechanism comprises: a motor support, a chip rotating motor, an integrated microfluidic chip, and a heating film support, a heating film and a heat-conducting aluminum ring arranged on the outside of the motor support in sequence from bottom to top.

3. The plasma sample automated detection device according to claim 1, characterized in that: The magnetic rod operating mechanism includes: a moving motor, a moving motor slider, a moving motor lead screw, and a motor support frame, a magnetic rod, a magnetic rod sleeve, and a magnetic rod moving motor which are sequentially arranged on the moving motor slider, wherein: the slider is fixedly connected to the support frame, the motor is fixedly connected to the support frame, the lead screw rotates to drive the slider to move vertically, the support frame moves vertically, and the magnetic rod sleeve is clamped by a magnetic rod sleeve fixing ring to achieve the installation of the magnetic rod sleeve, and the lead screw rotates to control the magnetic rod sleeve to enter each chamber to adsorb and transfer magnetic beads; the lead screw repeatedly rotates clockwise and counterclockwise to drive the magnetic rod sleeve to move back and forth in the vertical direction to achieve the beating and mixing of the liquid, the bottom of the telescopic rod of the motor is fixedly connected to the magnetic rod, and the motor drives the magnetic rod to move vertically, and the upward movement to leave the magnetic rod sleeve is demagnetization, and the downward movement to push the magnetic rod sleeve out of the fixing ring is unloading of the magnetic rod sleeve.

4. The plasma sample automated detection device according to claim 1, characterized in that: The pipette operating mechanism comprises: a pipette moving motor pad fixedly arranged on the device base, and a pipette moving motor, a pipette moving motor lead screw, a pipette moving motor slider, a pipette and a pipette head arranged thereon in sequence.

5. The plasma sample automated detection device according to claim 1, characterized in that: The fluorescence detection mechanism includes: an optical path fixture, a collimating lens, a laser transmitter, an optical fiber connected to the collimating lens, a fluorescence receiver and a circuit board, which are arranged in sequence, wherein: after the incident laser triggers the fluorescence reaction of the reagent in the chip detection area, it is received by the collimating lens and transmitted to the fluorescence receiver through the optical fiber. In the fluorescence receiver, the light path passes through the filter and is received by the fluorescence sensor for subsequent analysis.

6. The plasma sample automated detection device according to claim 5, characterized in that: The fluorescence detection mechanism further comprises a thin film temperature sensor connected to the circuit board, and the thin film temperature sensor is arranged on the heating film of the chip action mechanism.

7. An automated detection method based on the system according to any one of claims 1 to 6, characterized in that: include: Sample pretreatment, pipetting amplification and liquid separation detection operations are performed by adding plasma samples and proteinase K into the chip lysis chamber, installing the chip to the predetermined position of the device, closing the device door, and clicking the start detection button on the touch screen; installing the magnetic rod sleeve according to the preset program, the motor drives the chip to rotate so that the magnetic bead pre-storage chamber is aligned with the magnetic rod sleeve, and the magnetic rod movement mechanism drives the magnetic rod / magnetic rod sleeve to enter the magnetic bead pre-storage chamber to absorb the magnetic beads, and after transferring to the lysis chamber, the magnetic rod is separated from the magnetic rod sleeve, and the plasma sample is lysed under the auxiliary beating action of the magnetic rod sleeve, and the sample nucleic acid and magnetic beads are combined; the magnetic rod enters the magnetic rod sleeve again, adsorbs the magnetic beads, and lifts it above the chip, and the motor drives the chip to rotate so that the washing chamber is aligned with the magnetic rod sleeve, and the magnetic rod adsorbs the magnetic beads and enters the washing chamber. After completing two washes in sequence and drying the magnetic beads, the magnetic beads are transferred to the elution chamber, the elution solution elutes the nucleic acid from the magnetic beads, and the magnetic beads are taken away from the elution chamber by a magnetic rod to obtain the nucleic acid elution solution. The diluent is transferred to the nucleic acid elution chamber by an automatic pipetting device, and after being fully mixed with the nucleic acid elution solution, the mixed solution is transferred to the mixing chamber on the outer ring of the chip. The chip is then rotated under the control of a motor, and the centrifugal force is used to drive the mixed solution to the liquid dispensing and metering chamber to complete the equal division metering of the liquid. The chip is then rotated at high speed to break the capillary valve, so that the liquid in the metering chamber enters each reaction chamber. Under the action of the temperature control device, the nucleic acid is isothermally amplified in each reaction chamber. After the isothermal amplification is completed, the amplification product is detected by a fluorescence detection device to obtain the detection result.

8. The automated detection method according to claim 7, characterized in that: The sample pre-treatment specifically includes: Step 1: To avoid interference, move the slider up to the standby position to leave space; Step 2, after manually adding the plasma sample into the chamber, the mating ring of the chip is mounted on the motor; Step 3: The chip moves clockwise until the chamber reaches the bottom of the magnetic bar. The support frame moves downward, and after the magnetic bar cover is installed, the support frame moves upward to the standby position. Step 4: The chip rotates clockwise until the chamber reaches the bottom of the magnetic bar. The support frame moves down, and after the magnetic bar absorbs the magnetic beads, the support frame moves up to the standby position. Step 5: The chip rotates clockwise until the chamber reaches the bottom of the magnetic bar. At this time, the chamber contains the lysed sample. The support frame moves down, and the magnetic bar moves up to demagnetize. Then the magnetic bar sleeve begins to oscillate up and down, and the motor drives the chip to rotate clockwise and reversely in a small amplitude, simulating beating and mixing. This process is repeated five times. Step 6: Move the magnetic bar down, return to the inside of the magnetic bar sleeve, and re-absorb the magnetic beads. Step 7: Move the support frame upward and wait. Step 8: The chip moves clockwise, and the first washing chamber is in place; the magnetic rod sleeve moves downward, moves upward and returns after washing. Step 9: The chip moves clockwise, and the second washing chamber is in place; the magnetic rod sleeve moves downward, moves upward and returns after washing. Step 10: The chip moves clockwise and the elution chamber is in place; the magnetic rod cover moves downward and the bottom enters the elution solution and is immersed; after the elution operation is completed, the magnetic rod cover moves up and returns to the standby position. Step 11, the chip moves clockwise to place the chamber of the magnetic rod sleeve in place; the magnetic rod sleeve moves downward, the magnetic rod pushes out of the magnetic rod sleeve for unloading, and then the support frame moves up to the standby position.

9. The automated detection method according to claim 7, characterized in that: The pipetting amplification specifically comprises: Step a, pipette initialization, Step b: Rotate the chip clockwise and move the chamber to the position directly below the pipette. Remove the pipette tip from under the pipette and perform tip detection. Step c, the pipette moves up to a standby position; Step d, the chip rotates clockwise, and the chamber moves to the position directly below the pipette; the pipette moves downward, absorbs the dilution in the chamber, and then moves upward to the standby position; Step e: the chip rotates clockwise, and the chamber moves to the position directly below the pipette; the pipette moves downward to discharge the liquid, and then moves upward to the standby position; Step f, the chip is shaken back and forth, and the sample and the diluent in the chamber are mixed; Step g: the pipette moves down and aspirates the liquid, and then moves up to the standby position. Step h, the chip rotates clockwise, and the chamber moves to the position directly below the pipette; the pipette moves downward and adds the target sample from the sample adding hole into the chamber, and then moves upward to the standby position; Step i: The chip rotates clockwise and the chamber moves to the position directly below the pipette; the pipette performs a tip withdrawal operation.

10. The automated detection method according to claim 7, characterized in that: The liquid separation detection specifically includes: Step i, the heating film works, the heat-conducting aluminum ring heats up, and the temperature sensor is used for temperature feedback. The heating film starts and stops to control the heat-conducting aluminum ring to be in a constant temperature state, ensuring that the liquid in the outer ring chamber of the chip is in a constant temperature environment of 37±0.5℃; Step ii, the motor rotates slowly back and forth to shake and mix the liquid in the chamber; Step iii, reverse the valve to break the liquid into the siphon valve; then slowly accelerate to the liquid separation speed, and the liquid evenly enters the array liquid separation tank; Step iv, continue to increase the speed, and the liquid enters the array fluorescence detection area through the capillary valve for fluorescence amplification; Step v, rotate the chip in sequence to detect the fluorescence reaction of each chamber in the array fluorescence detection area. The fluorescence is received by the collimating lens and transmitted through the optical fiber. After passing through the filter in the fluorescence receiver, the signal is collected by the fluorescence sensor and the signal is returned to the circuit board to determine the positive and negative.

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