Plasma sample automatic detection device and method based on integrated microfluidic chip

Abstract

The application discloses a kind of based on integrated microfluidic chip's plasma sample automatic detection device and method, including: chip action mechanism, magnetic bar operating mechanism, pipette operating mechanism and fluorescence detection mechanism being arranged in device base, wherein: magnetic bar operating mechanism, pipette operating mechanism and fluorescence detection mechanism are respectively opposite the top and side of chip action mechanism.This application can greatly simplify equipment, reduce operation complexity, shorten detection time by the synergic control of chip and magnetic bar / magnetic bar sleeve motion mechanism, displacement mechanism, realize the super-multiplex detection of more than 30 biological targets in one-time detection.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological detection, and particularly relates to a plasma sample automatic detection device and method based on an integrated microfluidic chip. BACKGROUND

[0002] The current nucleic acid detection technology has a long process, complex operation, large volume of related equipment, high cost, and most of sample processing, nucleic acid extraction, purification, amplification and detection equipment are independent devices, the sample needs to be transferred between multiple devices, which is easy to cause pollution, and the number of detection target points is small, thereby limiting the application scene. SUMMARY

[0003] In view of the above problems in the prior art, the present application provides a plasma sample automatic detection device and method based on an integrated microfluidic chip, which can greatly simplify the equipment, reduce the operation complexity, shorten the detection time, and realize the super-multiplex detection of more than 30 biological targets at a time through the cooperative control of the chip, the magnetic rod / magnetic rod sleeve movement mechanism and the pipette mechanism.

[0004] The present application is achieved by the following technical solutions:

[0005] The present application relates to a plasma sample automatic detection device, which comprises a chip action mechanism, a magnetic rod operation mechanism, a pipette gun operation mechanism and a fluorescence detection mechanism arranged in the device base, wherein the magnetic rod operation mechanism, the pipette gun operation mechanism and the fluorescence detection mechanism are respectively opposite to the top and side of the chip action mechanism.

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

[0007] The integrated microfluidic chip comprises a motor matching ring in the center, a pipette head pre-storage chamber arranged on the bottom surface of the integrated microfluidic chip and located at the periphery, a magnetic rod sleeve pre-storage chamber, a magnetic bead liquid pre-storage chamber, a lysis liquid pre-storage chamber, two washing liquid pre-storage chambers, an elution liquid pre-storage chamber, a dilution liquid pre-storage chamber, and a sample adding 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 distribution groove, a capillary valve and a reaction chamber, wherein the motor is connected with the motor matching ring to drive the chip to rotate, so that each chamber can be aligned with the magnetic rod and the pipette gun, i.e. coaxial in the vertical direction.

[0008] The fluorescence detection mechanism comprises, in sequence, a light path fixing member, a collimating lens, a laser emitter, an optical fiber connected with 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, the fluorescence is received by the collimating lens and transmitted to the fluorescence receiver through the optical fiber. In the fluorescence receiver, the light path is received by the fluorescence sensor after passing through the optical filter, and subsequent analysis is performed.

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

[0010] Figure 1 is a schematic view of the present application;

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

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

[0013] Figure 4 is a schematic view of the magnetic rod / magnetic rod sleeve movement mechanism of the present application;

[0014] Figure 5 is a schematic view of the fluorescence detection mechanism of the present application;

[0015] In the figure: device base 1, chip 2, heating film 3, heating film support 401, heat-conducting aluminum ring 402, motor support 5, chip rotation motor 6, support frame moving motor 7, support frame moving motor sliding block 8, support frame moving motor lead screw 9, motor support 10, magnetic rod 11, magnetic rod sleeve 12, magnetic rod moving motor 13, pipette moving motor 14, pipette moving motor sliding block 15, pipette moving motor lead screw 16, pipette head 17, pipette 18, pipette moving motor pad 19, light path fixing member 20, collimating lens 21, optical fiber 22, laser emitter 23, fluorescence receiver 24, circuit board 25, fixing screw 26, thin film temperature sensor 27. DETAILED DESCRIPTION

[0016] As Figure 1 shown, the present embodiment relates to a kind of automatic detection device of plasma sample, comprising: chip action mechanism, magnetic rod operation mechanism, pipette operation mechanism and fluorescence detection mechanism being arranged in device base 1, wherein: magnetic rod operation mechanism, pipette operation mechanism and fluorescence detection mechanism are respectively opposite the top and side of chip action mechanism.

[0017] The chip action mechanism comprises: a motor support 5, a chip rotating motor 6, an integrated micro-fluidic chip 2, a heating film support 401, a heating film 3 and a heat-conducting aluminum ring 402 arranged in sequence from bottom to top, wherein the top of the heat-conducting aluminum ring 402 is in contact with the bottom of the periphery of the integrated micro-fluidic chip 2.

[0018] As shown in Figure 2 and Figure 3 , the integrated micro-fluidic chip 2 comprises: a motor matching ring 214 in the center, a pipette tip pre-storage chamber 201 arranged on the bottom surface of the integrated micro-fluidic chip 2 and located at the periphery thereof, a magnetic rod sleeve pre-storage chamber 202, a magnetic bead liquid pre-storage chamber 203, a lysis liquid pre-storage chamber 204, two washing liquid pre-storage chambers 205 and 206, an elution liquid pre-storage chamber 207, a dilution liquid pre-storage chamber 208, a sample adding area 209 arranged on the top surface of the integrated micro-fluidic 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 distribution groove 211, a capillary valve 215 and a reaction chamber 212, wherein the motor 6 is connected with 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, i.e. coaxial in the vertical direction.

[0019] As shown in Figure 1 and Figure 4 , the magnetic rod operation mechanism comprises: a moving motor 7, a moving motor sliding block 8, a moving motor lead screw 9, a motor support 10, a magnetic rod 11, a magnetic rod sleeve 12, a magnetic rod moving motor 13 arranged in sequence on the moving motor sliding block 8, wherein the sliding block 8 is fixedly connected with the support 10, and the motor 19 is fixedly connected with the support 10. The rotation of the lead screw 9 drives the vertical movement of the sliding block 8, and the vertical movement of the support 10, and the installation of the magnetic rod sleeve is realized by clamping the inner side of the magnetic rod sleeve 12 through the magnetic rod sleeve fixing ring 1001. The rotation of the lead screw 9 can control the magnetic rod sleeve 11 to enter each chamber to adsorb 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, thereby realizing the beating and mixing of the liquid. The bottom of the telescopic rod of the motor 19 is fixedly connected with the magnetic rod 11, and the motor 19 works to drive the magnetic rod 11 to move vertically, and the upward movement of the magnetic rod 11 away from the magnetic rod sleeve 12 is the demagnetization, and the downward movement of the magnetic rod 11 to push out the magnetic rod sleeve 12 from the fixing ring 1001 is the unloading of the magnetic rod sleeve.

[0020] The pipette operation mechanism comprises: a pipette moving motor cushion 19 fixedly arranged on the device base 1, and a pipette moving motor 14, a pipette moving motor lead screw 16, a pipette moving motor sliding block 15, a pipette 18 and a pipette tip 17 arranged in sequence thereon.

[0021] As shown in Figure 1 and Figure 5As shown, the fluorescence detection mechanism includes: an optical path fixture 20, a collimating lens 21, a laser emitter 23, an optical fiber 22 connected to the collimating lens 21, a fluorescence receiver 24, and a circuit board 25. 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. 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-mentioned system, which comprises the following steps: 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 by a motor so that the magnetic bead pre-storage chamber is aligned with the magnetic rod sleeve; a magnetic rod motion mechanism drives the magnetic rod / magnetic rod sleeve into the magnetic bead pre-storage chamber to absorb magnetic beads, and after transferring the magnetic rod 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; 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. The magnetic beads are washed twice in the elution chamber, and after drying, the magnetic beads are transferred to the elution chamber. The eluent elutes the nucleic acid from the magnetic beads. The magnetic beads are taken away from the elution chamber by a magnetic rod to obtain the nucleic acid eluent. The diluent is transferred to the nucleic acid elution chamber by an automatic pipetting device. After being fully mixed with the nucleic acid eluent, the mixed liquid is transferred to the mixing chamber on the outer circle of the chip. The chip is then rotated under the control of the motor, and the centrifugal force is used to drive the mixed liquid to the liquid separation and metering chamber to complete the equal division and 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.

[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 mounted on the motor 6 .

[0027] Step 103, the chip 2 moves clockwise, the chamber 202 reaches directly below the magnetic bar 11; the support frame 10 moves downward, installs the magnetic bar sleeve 12, and then moves upward to the standby position.

[0028] Step 104, the chip 2 rotates clockwise, the chamber 203 reaches directly below the magnetic bar 11; the support frame 10 moves downward, the magnetic bar 11 adsorbs the magnetic beads, and then the support frame 10 moves upward to the standby position.

[0029] Step 105, the chip 2 rotates clockwise, the chamber 204 reaches directly below the magnetic bar 11, and the chamber 204 contains the lysed sample at this time; the support frame 10 moves downward, and the magnetic bar 11 moves upward to demagnetize; then the magnetic bar sleeve 12 starts to move up and down in a reciprocating motion, and at the same time the motor 6 drives the chip 2 to rotate clockwise and counterclockwise with a small amplitude, simulating a beating mixing, for five times.

[0030] Step 106, the magnetic bar 11 moves downward to return to the inside of the magnetic bar sleeve 12 and re-adsorb the magnetic beads.

[0031] Step 107, the support frame 10 moves upward to standby.

[0032] Step 108, the chip 2 moves clockwise, the first washing chamber 205 is in place; the magnetic bar sleeve 12 moves downward, performs washing, and then moves upward to return.

[0033] Step 109, the chip 2 moves clockwise, the second washing chamber 206 is in place; the magnetic bar sleeve 12 moves downward, performs washing, and then moves upward to return.

[0034] Step 110, the chip 2 moves clockwise, the elution chamber 207 is in place; the magnetic bar sleeve 12 moves downward, the bottom enters the elution liquid and is immersed; after the elution operation is completed, the magnetic bar sleeve 12 moves upward to the standby position.

[0035] Step 111, the chip moves clockwise, the chamber 202 for placing the magnetic bar sleeve is in place; the magnetic bar sleeve 12 moves downward, the magnetic bar 11 pushes out the magnetic bar sleeve 12 to unload, and then the support frame 10 moves upward to the standby position.

[0036] The pipetting amplification specifically includes:

[0037] Step 201, the pipette is initialized.

[0038] Step 202, the chip 2 rotates clockwise, the chamber 201 moves to directly below the pipette 18; the pipette 18 moves downward to take a tip, and performs tip detection.

[0039] Step 203, the pipette 18 moves upward to the standby position.

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

[0041] Step 205, the chip 2 rotates clockwise, the chamber 207 moves to the position directly below the pipette 18; the pipette 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, the sample in the chamber 207 mixes with the diluent.

[0043] Step 207, the pipette 18 moves downward to suck the liquid and then moves upward to the standby position.

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

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

[0046] The liquid separation detection specifically includes:

[0047] Step 301, the heating film 3 works, the heat-conducting aluminum ring 402 is heated, 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, so that the liquid in the outer chamber of the chip 2 is in a 37±0.5℃ constant temperature environment.

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

[0049] Step 303, the valve is reversed, the liquid enters the siphon valve 210; then the speed is slowly increased to the liquid separation speed, and the liquid uniformly enters the array liquid separation groove 211.

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

[0051] Step 305, the chip 2 is rotated in sequence, and the fluorescence reaction of each chamber of the array fluorescence detection area 212 is detected. The fluorescence is received by the collimating lens and transmitted through the optical fiber, passes through the filter in the fluorescence receiver 24, and then the signal is collected by the fluorescence sensor, and the signal returns to the circuit board to determine the positive and negative.

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

[0053]

[0054] Compared with the existing technology, the integrated microfluidic chip involved in the present invention has the center of the inner circle chamber and the center of the sample addition hole of the outer circle sample addition chamber located on a concentric circle. The number of chip reaction chambers exceeds 30. Through the coordinated control of the chip and the magnetic rod / magnetic rod sleeve movement mechanism and the pipetting mechanism, it can greatly simplify the equipment, reduce the complexity of operation, shorten the detection time, and realize ultra-multiple detection of more than 30 biological targets at a time.

[0055] The above-mentioned specific implementation can be partially adjusted in different ways by those skilled in the art without departing from the principles and purpose of the present invention. The scope of protection of the present invention shall be based on the claims and shall not be limited by the above-mentioned specific implementation. All implementation schemes 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 and the pipette operation mechanism are respectively opposite to the top of the chip action mechanism, and the fluorescence detection mechanism is opposite to the side of the chip action mechanism; 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; The integrated microfluidic chip comprises: a motor mating ring located in the center; a pipette tip pre-storage chamber, a magnetic rod sleeve pre-storage chamber, a magnetic bead solution pre-storage chamber, a lysis solution pre-storage chamber, two wash solution pre-storage chambers, an eluent pre-storage chamber, a diluent pre-storage chamber, and a sample loading area, a siphon valve, a plurality of parallel test units, and a waste liquid chamber located on the top surface of the integrated microfluidic chip; each test unit comprises an array liquid separation tank, a capillary valve, and a reaction chamber; and a motor connected to the motor mating ring drives the chip to rotate so that each chamber can align with the magnetic rod and the pipette, i.e., be coaxial in the vertical direction. The fluorescence detection mechanism includes: an optical path fixture, a collimating lens, a laser emitter, an optical fiber connected to the collimating lens, a fluorescence receiver, and a circuit board, which are arranged in sequence. The incident laser triggers the fluorescence reaction of the reagent in the chip detection area, which is then received by the collimating lens and transmitted to the fluorescence receiver via the optical fiber. In the fluorescence receiver, the light passes through a filter and is received by a fluorescence sensor for subsequent analysis. The fluorescence detection mechanism further includes 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.

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

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

4. An automated detection method based on the automated detection device for plasma samples according to any one of claims 1 to 3, characterized in that: include: Sample pretreatment, pipetting amplification and liquid separation detection operations are performed by adding plasma samples and proteinase K to 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 and the magnetic rod sleeve into 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 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. The magnetic beads are washed twice in the elution chamber, and after drying, the magnetic beads are transferred to the elution chamber. The eluent elutes the nucleic acid from the magnetic beads. The magnetic beads are taken away from the elution chamber by a magnetic rod to obtain the nucleic acid eluent. The diluent is transferred to the nucleic acid elution chamber by an automatic pipetting device. After being fully mixed with the nucleic acid eluent, the mixed liquid is transferred to the sample addition area on the outer circle of the chip. The chip is then rotated under the control of the motor, and the centrifugal force is used to drive the mixed liquid to the liquid separation tank to complete the equal division measurement of the liquid. The chip is then rotated at high speed to break the capillary valve, so that the liquid in the liquid separation tank 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 test result.

5. The automated detection method according to claim 4, wherein: 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 to the lysate pre-storage chamber, the matching ring of the chip is installed on the motor; Step 3: The chip moves clockwise, and the magnetic rod sleeve pre-storage chamber reaches directly below the magnetic rod; the support frame moves downward, and after the magnetic rod sleeve is installed, the support frame moves up to the standby position; Step 4: The chip rotates clockwise, and the magnetic bead liquid pre-storage chamber reaches the bottom of the magnetic rod; the support frame moves down, and after the magnetic rod absorbs the magnetic beads, the support frame moves up to the standby position. Step 5: The chip rotates clockwise, and the lysis solution pre-storage chamber reaches the bottom of the magnetic rod. At this time, the lysis solution pre-storage chamber contains the lysed sample; the support frame moves down, and the magnetic rod moves up and demagnetizes; then the magnetic rod sleeve begins to oscillate up and down, and the motor drives the chip to rotate clockwise and counterclockwise in a small amplitude, simulating beating and mixing, and continues for five times. Step 6: Move the magnetic rod down, return to the inside of the magnetic rod sleeve, and re-absorb the magnetic beads. Step 7: Move the support frame upwards and wait for the next step. Step 8: The chip moves clockwise, and the first washing solution pre-storage chamber is in place; the magnetic rod sleeve moves downward, washes, and then moves back upward. Step 9: The chip moves clockwise, and the second washing solution pre-storage chamber is in place; the magnetic rod sleeve moves downward, washes, and then moves back upward. Step 10: The chip moves clockwise, and the eluent pre-storage chamber is in place; the magnetic rod sleeve moves downward, and the bottom enters the eluent and is immersed; after the elution operation is completed, the magnetic rod sleeve moves up and returns to the standby position. Step 11: The chip moves clockwise, and the magnetic rod sleeve pre-storage chamber for placing the magnetic rod sleeve is in place; the magnetic rod sleeve moves downward, and the magnetic rod pushes out of the magnetic rod sleeve for unloading, and then the support frame moves up and returns to the standby position.

6. The automated detection method according to claim 4, wherein: The pipetting amplification specifically includes: Step a, pipette initialization, Step b: Rotate the chip clockwise, move the pre-storage chamber of the pipette tip to the bottom of the pipette; remove the pipette tip from under the pipette and perform pipette tip inspection. Step c, the pipette moves up to the standby position; Step d: The chip rotates clockwise, and the diluent pre-storage chamber moves to the position directly below the pipette; the pipette moves downward, draws the diluent from the diluent pre-storage chamber, and then moves upward to the standby position; Step e: The chip rotates clockwise, and the eluent pre-storage 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 to mix the sample and the diluent in the eluent pre-stored chamber; Step g: Move the pipette downward to aspirate the liquid, then move it upward to the standby position. Step h: The chip rotates clockwise, and the sample loading chamber moves to the position directly below the pipette; the pipette moves downward and adds the target sample from the loading hole to the sample loading chamber, and then moves upward to the standby position; Step i: The chip rotates clockwise, and the pre-storage chamber of the pipette tip moves to the bottom of the pipette; the pipette performs the operation of retracting the pipette tip.

7. The automated detection method according to claim 4, wherein: The liquid separation detection specifically includes: Step i: The heating film works, the heat-conducting aluminum ring heats up, and the temperature sensor provides temperature feedback. The heating film starts and stops to control the heat-conducting aluminum ring to maintain a constant temperature, ensuring that the liquid in the outer reaction chamber of the chip is kept at a constant temperature of 37±0.5°C. Step ii: The motor rotates slowly back and forth to shake and mix the liquid in the sample adding area 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 separation tank; Step iv: Continue to increase the speed, and the liquid enters the reaction chamber through the capillary valve for fluorescence amplification; Step v: Rotate the chip in sequence to detect the fluorescence reaction of each reaction chamber. The fluorescence is received by the collimating lens and transmitted through the optical fiber. After passing through the filter in the fluorescence receiver, the fluorescence sensor collects the signal and the signal is returned to the circuit board to determine whether it is positive or negative.

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