Detection method for hs-TnI for non-disease diagnosis purposes based on quantum dot microspheres

Abstract

The present invention discloses a hs-TnI detection method based on quantum dot microspheres, which directly reacts a sample with a first solution containing antibody 1 and a second solution containing antibody 2 to generate a sandwich structure complex of antibody 1-antigen-antibody 2, and the antigen and antibody react in a reaction cup, incubate at a constant temperature, and the reaction time is longer, and the reaction is more sufficient, which is conducive to the improvement of detection sensitivity. At the same time, during chromatography, with the high affinity of streptavidin and biotin, the complex obtained by the reaction can be better captured on the T line, so that the reaction can be made more sensitive, and the detection time is also faster relative to the existing reagent strips. Further, combined with the advantages of high sensitivity and wide linear range of quantum dot microspheres, detection repeatability (CV) is better, CV is increased from the traditional 15wt% to within 8wt%, and the detection sensitivity is as high as 2pg / ml, while the linear range is 2-30000pg / ml, and full coverage is achieved for low-concentration and high-concentration samples.

Description

Technical Field

[0001] The present invention relates to the technical field of immunoassay detection, in particular to a detection method for hs-TnI for non-disease diagnosis purposes based on quantum dot microspheres. Background Art

[0002] High-sensitivity troponin I (hs-TnI) is a specific protein that reflects myocardial infarction, so its detection is necessary.

[0003] Fluorescence immunoassay is a commonly used detection method, and detection using fluorescent immunochromatographic reagent strips is an available method. The invention patent with authorization announcement number CN102520192B discloses a fluorescent immunochromatographic kit for quantitative detection of troponin I / creatine kinase isoenzymes / myoglobin.

[0004] The minimum detection limit of this fluorescent immunochromatography kit and its detection method is 0.1 ng / mL, and the coverage range is not low enough. At the same time, its detection time is not less than 13 minutes, which is time-consuming. Summary of the invention

[0005] The purpose of the present invention is to solve the above problems existing in the prior art and to provide a detection method for hs-TnI based on quantum dot microspheres for non-disease diagnosis purposes.

[0006] The purpose of the present invention is achieved through the following technical solutions:

[0007] The non-disease diagnosis purpose detection method of hs-TnI based on quantum dot microspheres comprises the following steps:

[0008] S1, preparing a reagent strip, wherein the reagent strip comprises a sample pad, an NC detection membrane, and absorbent paper which are sequentially arranged on a substrate, wherein the T line of the NC detection membrane is fixed with streptavidin, and the C line is fixed with mouse anti-DNP monoclonal antibody;

[0009] S2, preparing a first solution and a second solution, wherein the first solution comprises quantum dot microspheres coupled to troponin antibody 1 and quantum dot microspheres coupled to DNP-BSA, and the second solution comprises troponin antibody 2;

[0010] S3, mixing a certain sample, the first solution and the second solution and reacting them for a certain period of time to obtain a reaction solution;

[0011] S4, taking a certain amount of reaction solution and dropping it into the sample addition hole of the reagent strip, after chromatography for a certain period of time, the T line and C line of the reagent strip are detected by a quantum dot detector to obtain the detection result.

[0012] Preferably, the preparation process of the first solution is as follows:

[0013] S21, preparing a first quantum dot microsphere solution including quantum dot microspheres coupled to troponin antibody 1;

[0014] S22, preparing a second quantum dot microsphere solution including quantum dot microspheres coupled with DNP-BSA;

[0015] S23, diluting the first quantum dot microsphere solution and the second quantum dot microsphere solution respectively with microsphere buffer and mixing them to obtain a first solution.

[0016] Preferably, the blocking agent used in the preparation process of the first quantum dot microsphere solution and the second quantum dot microsphere solution is CE510 blocking agent.

[0017] Preferably, the particle size of the quantum dot microspheres is greater than or equal to 10 nanometers and less than 100 nanometers.

[0018] Preferably, the OD of the first solution is 340 Between 0.10-0.20.

[0019] Preferably, the microsphere buffer comprises sucrose and Tween 20.

[0020] Preferably, the troponin antibody 2 is conjugated to biotin.

[0021] Preferably, in S3, the sample, the first solution and the second solution are allowed to react at a constant temperature for 3-5 minutes.

[0022] Preferably, in S4, the chromatography time is 4-6 minutes.

[0023] Preferably, S3 and S4 are implemented by an automated detection device.

[0024] The advantages of the technical solution of the present invention are mainly reflected in:

[0025] The method of the present invention allows the sample to directly react with the first solution containing antibody 1 and the second solution containing antibody 2 to generate a sandwich structure complex of antibody 1-antigen-antibody 2. The antigen and antibody react in the reaction cup, incubate at a constant temperature, and the reaction time is longer, the reaction is more complete, and it is conducive to the improvement of detection sensitivity. At the same time, during chromatography, with the help of the high affinity of streptavidin, the complex obtained by the reaction can be better captured on the T line, so that the reaction can be more sensitive, and the detection time is also faster than that of the existing reagent strip. Further, combined with the advantages of high sensitivity and wide linear range of quantum dot microspheres, and the reagent strip has no release pad, there is no problem of uneven drying and incomplete release of quantum dot microspheres on the release pad, and the detection repeatability (CV) is better, CV is increased from the traditional 15wt% to within 8wt%, and the detection sensitivity is as high as 2pg / ml, while the linear range is 2-30000pg / ml, and low-concentration and high-concentration samples are fully covered, and full-scale high-dispersion quantum dot immunoassay is truly achieved.

[0026] The present invention improves the dispersibility of quantum dot microspheres by selecting the particle size of quantum dot microspheres, the configuration process of the first solution and the design of the microsphere buffer solution, which is beneficial to improving the accuracy of subsequent detection results.

[0027] In the reagent strip of the present invention, the detection antibody coupled with biotin is combined with streptavidin, and after biotin-avidin amplification, the detection background is reduced, the signal is increased several times, and the detection signal level is greatly improved, so that samples with lower concentrations can be detected, and the detection sensitivity is improved.

[0028] The main detection process of the detection method of the present invention can be implemented by automated equipment, with low operating difficulty, and can realize parallel detection of multiple reagent strips with high speed and efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 It is a result schematic diagram of the linear range test of the present invention;

[0030] Figure 2 is a stereogram of the automated testing equipment of the present invention;

[0031] Figure 3 is a top view of the automated testing equipment of the present invention;

[0032] Figure 4 is a side view of the automated testing device of the present invention;

[0033] Figure 5 It is a first stereoscopic view of the automated testing device of the present invention with the carrier plate and a testing mechanism hidden.

[0034] Figure 6It is a second stereoscopic view of the automated testing device of the present invention without the carrier plate and a testing mechanism.

[0035] Figure 7 is a three-dimensional diagram of a reaction turntable of the present invention;

[0036] Figure 8 is a partial cross-sectional view of the reaction turntable portion of the present invention;

[0037] Fig. 9 is a three-dimensional diagram of the carrier plate and the structure thereon of the present invention;

[0038] Fig.10 is a side view of the rotating structure of the carrier plate of the present invention;

[0039] Fig.11 yes Fig.10 Bottom view of

[0040] Fig.12 is a perspective view of a liquid adding device of the present invention;

[0041] Fig.13 It is a three-dimensional diagram of the carrier plate of the present invention and the cooperation between the liquid adding device and the pressing device thereon;

[0042] Fig.14 It is a three-dimensional diagram of the liquid adding device of the present invention having a shaking mechanism;

[0043] Fig.15 It is a three-dimensional diagram of the feeding mechanism of the present invention;

[0044] Fig.16 is a cross-sectional view of the feeding mechanism of the present invention;

[0045] Fig.17 is a side view of the feeding mechanism of the present invention;

[0046] Fig.18 yes Fig.15 A partial enlarged view of the

[0047] Fig.19 is a top view of the feeding mechanism of the present invention;

[0048] Fig. 20 is a cross-sectional view of an injection assembly of the present invention;

[0049] Fig.21 is a partial stereoscopic diagram of the liquid transfer mechanism of the present invention;

[0050] Fig. 22 It is a side view of the push-pull mechanism and the detection mechanism of the present invention;

[0051] Fig.23 is a cross-sectional view of the push-pull mechanism and the detection mechanism of the present invention;

[0052] Fig.24 It is an end view of the push-pull mechanism and the detection mechanism of the present invention. DETAILED DESCRIPTION

[0053] The purpose, advantages and features of the present invention will be illustrated and explained by the non-limiting description of the following preferred embodiments. These embodiments are only typical examples of the application of the technical solution of the present invention, and any technical solution formed by equivalent replacement or equivalent transformation falls within the scope of protection claimed by the present invention.

[0054] In the description of the scheme, it should be noted that the terms "center", "upper", "lower", "left", "right", "front", "back", "vertical", "horizontal", "inside", "outside" and the like indicate positions or positional relationships based on the positions or positional relationships shown in the drawings, and are only for the convenience and simplification of description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the present invention. In addition, the terms "first", "second", and "third" are used for descriptive purposes only, and cannot be understood as indicating or implying relative importance.

[0055] Example 1

[0056] The following is an explanation of the non-disease diagnosis purpose detection method of hs-TnI based on quantum dot microspheres disclosed in the present invention in conjunction with the accompanying drawings, which comprises the following steps:

[0057] S1, preparing a reagent strip, wherein the reagent strip comprises a sample pad, an NC detection membrane, and absorbent paper which are sequentially arranged on a substrate, wherein the T line of the NC detection membrane is fixed with streptavidin, and the C line is fixed with mouse anti-DNP monoclonal antibody;

[0058] S2, preparing a first solution and a second solution, wherein the first solution comprises quantum dot microspheres coupled to troponin antibody 1 and quantum dot microspheres coupled to DNP-BSA, and the second solution comprises troponin antibody 2;

[0059] S3, mixing a certain sample, the first solution and the second solution and reacting them for a certain period of time to obtain a reaction solution;

[0060] S4, taking a certain amount of reaction solution and dropping it into the sample addition hole of the reagent strip, after chromatography for a certain period of time, the T line and C line of the reagent strip are detected by a quantum dot detector to obtain the detection result.

[0061] Specifically, the preparation of the reagent strip includes the main processes of preparing the NC detection film, preparing the sample pad, assembling the reagent strip, etc.

[0062] (1) The preparation process of NC detection membrane is as follows:

[0063] S11, dilute streptavidin (manufacturer: Feipeng Biotechnology Co., Ltd., brand: SA-AG-E1-003) in a streaking buffer to obtain a T-line solution, wherein the concentration of the T-line solution is between 0.5-1 mg / ml; the streaking buffer includes 20 mM PBS buffer, 2 wt% trehalose, and 0.05 wt% Proclin300.

[0064] S12, dilute mouse anti-DNP monoclonal antibody (manufacturer: Xiamen Tongrenxin Biotechnology Co., Ltd., brand: ZKC21) in the streaking buffer to obtain a C-line solution, wherein the concentration of the C-line solution is between 1-2 mg / ml.

[0065] S13, using a film stripper, such as a film stripper from Biodot, to stripe the T-line solution and the C-line solution on the NC film, and drying to obtain the NC detection film.

[0066] (2) The preparation process of the sample pad is as follows:

[0067] A sample pad buffer was prepared, wherein 0.6 wt % Tween 20, 0.4 wt % BSA, and 0.05 wt % Proclin 300 were dissolved in 50 mM PB buffer at pH 6.0.

[0068] Soak the RB65 glass fiber in the sample pad buffer solution, take it out after five minutes, place it in a drying room (humidity <40wt%) to dry, and then cut it into the required size to obtain the sample pad.

[0069] (3) The assembly process of the reagent strip is as follows:

[0070] A sample pad, an NC detection film, and absorbent paper are respectively pasted on a substrate, wherein the substrate is a PVC rubber board. Meanwhile, the NC detection film is located between the sample pad and the absorbent paper. Meanwhile, one end of the sample pad can be overlapped on one end of the NC detection film, and the absorbent paper can be overlapped on the other end of the NC detection film. The substrate with the sample pad, the NC detection film, and the absorbent paper attached is cut and loaded into a shell to obtain the reagent strip. A reaction cup, a sample loading hole corresponding to the sample pad, and an observation port corresponding to the NC detection film are formed on the shell.

[0071] In S2, the preparation process of the first solution is as follows:

[0072] S21, preparing a first quantum dot microsphere solution including quantum dot microspheres coupled to troponin antibody 1;

[0073] Specifically, the quantum dot microspheres with a particle size greater than or equal to 10 nanometers and less than 100 nanometers developed by Kundao Bio are diluted with MES buffer at pH 6.1 and centrifuged to remove the supernatant, and the precipitate is ultrasonically re-dissolved using the above-mentioned MES buffer; EDC and NHS are added to activate the carboxyl groups of the quantum dot microspheres for 30 minutes, wherein the mass ratio of EDC to NSH is 1:5, and the mass ratio of EDC to quantum dot microspheres is 1:1250-2500; the obtained solution is centrifuged to remove the supernatant, and after ultrasonic re-dissolution using the above-mentioned MES buffer, troponin antibody 1 is added, and the mass ratio of quantum dot microspheres to troponin antibody 1 is between 1:0.1-10, and the reaction is carried out for 1 hour; after the reaction, a blocking agent is added to block for 2 hours, and the blocking agent is preferably a ce510 blocking agent; after the blocked solution is centrifuged to remove the supernatant, a storage buffer is added, and the solution is ultrasonically mixed, and the storage buffer is a known buffer and is not limited here.

[0074] S22, prepare a second quantum dot microsphere solution including quantum dot microspheres coupled with DNP-BSA (manufacturer: Xiamen Tongrenxin Biotechnology Co., Ltd., brand: ZKC11); the specific preparation process is the same as the process of S21, which will not be repeated here.

[0075] S23, diluting the first quantum dot microsphere solution and the second quantum dot microsphere solution with microsphere buffer respectively and mixing them to obtain a first solution. 340 The OD of the second quantum dot microsphere solution after dilution with the microsphere buffer is between 0.1 and 0.2. 340 Between 0.01-0.03; the OD of the first solution 340 The OD is between 0.10 and 0.20, preferably 0.15. 340 The microsphere buffer comprises 50 mM Tris buffer at pH 8.0, 0.9 wt% sodium chloride, 0.05 wt% Tween 20, 0.5 wt% bovine serum albumin, 1 wt% sucrose, and 0.05 wt% PC300.

[0076] Dispersion analysis of quantum dot microspheres:

[0077] The prepared first solution was divided into several portions and placed under the condition of 2-8 °C. The aggregation of the solution was detected by an Uncle analyzer in January, March, June, and December respectively. The specific results are shown in Table 1. Among them, the dispersion coefficient PDI (Polydispersity index) is a dimensionless value reflecting the width of the particle size distribution, ranging from 0 to 1. The smaller the value, the more uniform the particle size and the more concentrated the particle size distribution. PDI < 0.1: monodisperse, narrow distribution; 0.1 < PDI < 0.2: medium aggregation; PDI > 0.2: polydisperse, severe aggregation.

[0078] Table 1:

[0079]

[0080] The above results show that the process of the present invention can achieve high dispersibility of quantum dot microspheres in the first solution.

[0081] The reason for this effect is that by selecting quantum dot microspheres with smaller particle sizes, the quantum dot microspheres have better dispersibility. At the same time, the sucrose included in the microsphere buffer has a certain viscosity, which reduces the Brownian motion of the quantum dot microspheres in the solution, making the quantum dot microspheres not easily collide with each other. In addition, Tween 20 in the microsphere buffer is a non-ionic surfactant, which improves the dispersibility of the solution. Further, when the solution is sealed, a small molecule ce510 is added. Compared with macromolecules such as BSA, casein, and skim milk powder, the small molecule ce510 has a better sealing effect and can ensure the dispersion effect of the solution.

[0082] In S2, the preparation process of the second solution is as follows:

[0083] Using a known method, troponin antibody 2 was conjugated with biotin at a molar ratio of 1:10-20. The biotin is, for example, NHS ester. The conjugated biotin troponin antibody 2 was diluted with a biotin buffer to an antibody concentration of 0.5-1 μg / ml to obtain the second solution. The biotin buffer was obtained by dissolving 0.9 wt% sodium chloride, 0.05 wt% Tween 20, 0.5 wt% bovine serum albumin, 1 wt% sucrose, and 0.05 wt% PC300 into 50 mM, pH 8.0 Tris buffer.

[0084] In S3, 40-50 μl of the first solution and 40-50 μl of the second solution were added to a reaction cup of the reagent strip, preferably 45 μl. Then, 10-15 μl of the sample, preferably 10 μl of the sample, was added to the reaction cup and mixed. The reaction cup of the reagent strip was kept at a constant temperature in an environment of 30-35 °C for 3-5 minutes, and more preferably at a constant temperature in an environment of 31-32 °C for 5 minutes.

[0085] In S4, 80 microliters of the reaction solution is added to the sample well of the reagent strip, and the chromatography time is 4-6 minutes, preferably 5 minutes, so that the entire reaction time can be controlled within 10 minutes. Finally, the quantum dot fluorescence signals of the T line and C line on the test strip are tested by a quantum dot immunoassay to quantitatively detect the content of the analyte in the sample.

[0086] According to the above method, the following performance tests are performed:

[0087] (1) Minimum detection limit:

[0088] The above reagent strip, the first solution, and the second solution were used to detect the blank buffer solution of high-sensitivity troponin (repeated 20 times), and the (blank mean) and S (standard deviation), statistics" The concentration values ​​of ", the results are shown in Table 2 below:

[0089] Table 2:

[0090]

[0091] It can be seen from the above that the minimum detection limit is ≤2pg / mL.

[0092] (2) Linear range

[0093] Samples of different concentrations were mixed into 8 concentrations in proportion, and each concentration was tested 4 times. The mean value (Yi) of the test results was calculated respectively. The results are shown in Table 3 below. The linear regression equation was obtained with the dilution concentration (Xi) as the independent variable and the mean value (Yi) of the test results as the dependent variable, and the linear regression correlation coefficient (r2) was calculated. Figure 1 shown.

[0094] Table 3:

[0095]

[0096] It can be concluded from the above test results that: within the range of 2pg / ml-30000pg / ml, r2 is 1 and there are no outliers. Therefore, within the range of 2pg / ml-30000pg / ml, full coverage of low-concentration and high-concentration samples is achieved. Although 30000pg / ml is set as the upper detection limit in this embodiment, in other embodiments, other upper detection limits may be set, which may be higher, depending on the requirements for consistency of the standard curve.

[0097] (3) Precision-Repeatability

[0098] Samples with low, medium and high concentrations (20pg / mL, 150pg / mL, 1500pg / mL) were tested separately, and each concentration was tested 10 times. The mean (M) and standard deviation (S) of the 10 measurement results were calculated, and the coefficient of variation (CV) was calculated, CV=S / M×100wt%; the specific test results are shown in Table 4 below.

[0099] Table 4:

[0100]

[0101] It can be seen from the above table that the test results have high accuracy and consistency.

[0102] Example 2

[0103] S3 and S4 of the above detection method are preferably implemented by an automated detection device, such as the attached Figure 2 -Attached Figure 4 As shown, the automated detection device includes a base 100, on which are disposed:

[0104] A reaction turntable 200, which is rotatably disposed on the base 100 and connected to a first rotating mechanism for driving its rotation. The reaction turntable 200 is provided with radial slots 210 for inserting a reagent strip B radially therefrom;

[0105] The carrier plate 300 is concentric with the reaction turntable 200 and is arranged above the reaction turntable 200. The carrier plate 300 is provided with a liquid adding device 301 for adding the first solution and the second solution to the reagent strip B on the reaction turntable 200; the carrier plate 300 is provided with a first placement position for placing a test tube containing a sample and a second placement position for placing a tip box containing a tip;

[0106] A loading mechanism 400 is disposed on the outer periphery of the reaction turntable 200 and is used to insert the reagent strips B one by one into the radial slots 210 of the reaction turntable 200;

[0107] The pipetting mechanism 500 includes an injection assembly 501 and a sample adding drive mechanism 515 for driving the injection assembly 501 to move horizontally and vertically. The pipetting mechanism 500 is used to suck the sample from the test tube containing the sample at the first placement position and add it to the reaction cup B1 of the reagent strip B on the reaction turntable 200, and to suck the reaction liquid in the reaction cup B1 and add it to the sample adding hole of the reagent strip B.

[0108] The push-pull mechanism 600 is arranged on the outer periphery of the reaction turntable 200, and is used to enable the reagent strip on the reaction turntable 200 to be transferred from the radial slot to the detection mechanism and to be withdrawn from the detection mechanism;

[0109] The detection mechanism 700 is arranged beside the push-pull mechanism 600 and is used to read the reagent strip inserted thereon.

[0110] As attached Figure 5 As shown, the base 100 includes an upper plate 110 and a lower plate 120 which are distributed up and down and connected by a column. The bottom of the lower plate 120 is connected with a support foot and / or a universal wheel.

[0111] As attached Figure 5 -Attached Figure 8 As shown, the reaction turntable 200 includes an outer ring 201 and an inner ring 202 that are concentrically connected. The outer ring 201 is provided with a circle of radial grooves 203. The inner ring 202 includes reaction chambers 204 that correspond to and dock with each of the radial grooves 203. A heating film 205 is provided at the bottom of the reaction chamber 204 so that the reaction chamber can be kept at a constant temperature. A liquid adding hole 206 is provided at the top of the reaction chamber 204.

[0112] As attached Figure 7 As shown, the outer ring 201 includes a lower ring 207, and the lower ring 207 is a circular ring. The top surface of the lower ring 207 is provided with a circle of partitions 208 that are evenly distributed around the circumference and are roughly fan-shaped. The inner arc of the partition 208 faces the center of the reaction turntable 200, and the outer arc of the partition 208 faces outward. Each of the partitions 208 is provided with a shielding plate 209, and both sides of the shielding plate 209 extend outside the two sides of the partition 208. Two adjacent partitions 208 and the shielding plates 209 on them define a radial through groove 203. The inner end of each radial channel is connected to the lateral opening of a reaction chamber 204 on the inner ring, and a reaction chamber 204 and a radial through groove 203 form a radial slot 210. As shown in the attached Figure 8 As shown, the inner ring 202 is also a circular ring, and in order to facilitate power supply to the heating film 205 , a first slip ring 130 concentric with the reaction turntable 200 is provided on the upper plate 110 of the base.

[0113] The reaction turntable 200 may be disposed on the upper plate 110 via a hollow servo turntable.

[0114] As attached Figure 8As shown, the reaction turntable 200 is rotatably arranged on the base 100 through a group of support rollers 211 and the horizontal position is limited by a group of limiting rollers 212. The support rollers 211 are bearings and are rotatably arranged on the upper plate 110 through a horizontal axis mounted on the upper plate 110. The number of the support rollers 211 can be designed as needed, for example, 3-5, which are evenly distributed on the upper plate 110 and their axes intersect perpendicularly with the axis of the reaction turntable 200. The limiting rollers 212 are bearings and are arranged on the upper plate 110 through a vertical axis. The number of the limiting rollers 212 can also be designed as needed, preferably 3-5, which are evenly distributed on the circumference and attached to the inner side wall of the driving ring 213 below the lower ring 207.

[0115] As attached Figure 8 As shown, the first rotating mechanism includes a first driving wheel 214 that is tightly attached to the outer circumference of the reaction turntable 200, and the first driving wheel 214 is connected to a first motor 215 that drives it to rotate. Specifically, the first driving wheel 214 is attached to the outer circumference of the driving ring 213, and the first driving wheel 214 can be a gear, and correspondingly, the outer circumference of the driving ring 213 is a tooth groove that matches the gear. Of course, in other embodiments, the first driving wheel 214 can also be a rubber wheel. The first driving wheel 214 can be directly connected to the power output shaft of the first motor 215, and the first motor 215 can be a known reduction motor and fixed on the lower plate 120.

[0116] As attached Figure 2 -Attached Figure 4 And attached Fig. 9 As shown, the carrier plate 300 may be a circular ring-shaped body, the diameter of the inner hole of which is larger than the outer diameter of the inner ring 202 , and is disposed on the base and located above the reaction turntable 200 .

[0117] The structure of the liquid adding device 301 can be designed as required. For example, in one embodiment, the liquid adding device 301 includes at least one liquid storage tank, preferably a plurality of liquid storage tanks, and the plurality of liquid storage tanks are distributed in an arc shape on the carrier plate 300, and the arc shape they present is concentric with the carrier plate 300. In order to avoid using a pipetting mechanism to pipette the first solution and the second solution, a leakage hole can be provided at the bottom of each liquid storage tank, and a mass flow meter is provided at the leakage hole to control the on-off and flow rate of the leakage hole at each liquid storage tank, and an avoidance hole corresponding to the leakage hole is provided on the carrier plate.

[0118] As attached Fig.10 , Attachment Fig.11As shown, the carrier plate 300 can be arranged on the base 100 so as to rotate relative to the base 100 and is connected to a second rotating mechanism 305 that drives the carrier plate 300 to rotate. Specifically, the carrier plate 300 is arranged on a group of supporting rollers 303, and the horizontal position is limited by a group of limiting rollers 302. Each supporting roller 303 and a limiting roller 302 are arranged on a mounting seat 304, and the mounting seat 304 is fixed on the upper plate 110 in a distributed manner along the circumference of the carrier plate 300, and each limiting roller 302 is attached to the outer peripheral wall of the carrier plate 300. The structure of the second rotating mechanism 305 is the same as that of the first rotating mechanism, and will not be described here. The difference is that the driving motor of the second rotating mechanism 305 is arranged on the lower plate 120.

[0119] As attached Fig.11 , Attachment Fig.12 As shown, in a more preferred embodiment, in order to reduce the power source required by the liquid adding device 301 and avoid setting up multiple mass flow meters.

[0120] The liquid adding device includes a mounting block 306, and a group of limiting holes 308 for fixing a syringe 307 are provided at the mounting block 306. The syringe 307 is used to absorb the first solution and the second solution. The limiting holes 308 are arranged in an arc shape, and the arc shape of the limiting holes 308 is concentric with the carrier plate 300. The carrier plate 300 is provided with avoidance holes 309 corresponding to the limiting holes 308, so that the syringe 307 located at each of the limiting holes 308 can drip the first solution and the second solution into the reaction cup B1 of the reagent strip B on the reaction turntable 200 through the avoidance holes 309.

[0121] As attached Fig.12 As shown, in order to extend the service life of the first solution and the second solution at the mounting block 306, the first solution and the second solution at the mounting block 306 need to be refrigerated. Specifically, the side of the mounting block 306 is also connected to a refrigerator 310. At the same time, the top of the mounting block 306 is also hinged with an upper cover 311, and the upper cover 311 is detachably connected to the shield 312 on the outer periphery of the mounting block 306. The upper cover 311 is provided with a through hole corresponding to each of the limiting holes 308 for the syringe 307 to pass through. At the same time, the mounting block 306 and the shield 312 are also provided with an observation hole 313 that is connected to each of the limiting holes 308 and is located on the side of the limiting hole so as to intuitively determine the remaining amount of the first solution and the second solution in the syringe 307.

[0122] As attached Figure 6 , Attachment Fig.13As shown, in order to allow the syringe 307 in the limiting hole 308 to drip, a pressing device 800 for driving the piston core rod of the syringe 307 at the liquid adding device 301 to move downward is provided on the outer side of the carrier plate 300, and the pressing device 800 is close to the feeding mechanism. The pressing device 800 includes a pressure head 801 and a pressing drive mechanism 802 for driving the pressure head 801 to move up and down. The specific structure of the pressing drive mechanism 802 can be designed as needed, for example, it can be a linear motor, a hydraulic cylinder or an electric cylinder. In this embodiment, the pressing drive mechanism 802 includes a pressing drive motor and a lead screw driven by the motor to rotate. The nut of the lead screw is connected to the pressure head 801 through a transfer block, and the transfer block is slidably arranged on a slide rail. The carrier plate can make the syringes at the liquid adding device rotate to the bottom of the pressure head 801 in sequence.

[0123] As attached Fig.13 As shown, there are two liquid adding devices, and they are evenly arranged on the circumference of the carrier plate 300. There can also be two pressing devices, and they are evenly arranged on the circumference.

[0124] As attached Fig.14 As shown, in order to make the quantum dot microspheres of the first solution and the second solution in the syringe 307 disperse more evenly, the mounting block 306 is connected to a shaking mechanism 314 that drives it to at least reciprocate linearly. Specifically, the shaking mechanism 314 includes a swing arm 315 connected to the bottom of the mounting block 306, the swing arm 315 is connected to a slide rail assembly, the slide rail assembly includes a first slide groove 316 consistent with the extension direction of the swing arm, the first slide groove 316 is slidably arranged on a first horizontal rail 317, the first horizontal rail 317 extends along the length direction of the swing arm 315, and a second horizontal rail 318 is arranged below the first horizontal rail 317 in a cross-shaped manner, and the second horizontal rail 318 is slidably arranged on a second slide groove 319, and the second slide groove 319 is arranged on the carrier plate 300. One end of the swing arm 315 is eccentrically and pivotally connected to the crank 321 through the pivot shaft 320. The crank 321 is connected to the motor shaft of the shaking motor 322. The shaking motor 322 can be a reduction motor. The motor shaft is parallel to the pivot shaft 320. When the motor shaft drives the crank 321 to rotate, the swing arm 315 is driven to swing back and forth, thereby realizing the shaking of the mounting block 306.

[0125] As attached Fig. 9 , Attachment Fig.13As shown, the first placement position on the carrier plate 300 is used to place and limit the test tube rack 900, and the test tube rack 900 is used to store test tubes 910 containing samples. The first placement position includes two test tube rack placement grooves 323, and the two test tube rack placement grooves 323 are arc grooves concentric with the carrier plate 300. The test tubes 910 on the test tube rack 900 are also distributed in an arc shape concentric with the carrier plate 300. Furthermore, the test tubes 910 on the test tube rack 900 located in the test tube rack placement grooves and the limiting holes 308 can be located on the same arc.

[0126] As attached Fig. 9 , Attachment Fig.13 As shown, the second placement position on the carrier plate 300 is used to place and limit the suction head box A, and the suction head box A is used to store suction heads A1, which includes a plurality of slots extending in the vertical direction, and the suction heads A1 are inserted into the slots with the tip facing downward and the suction heads A1 protrude out of the slots. The second placement position includes two storage slots 324, and each storage slot 324 can limit a suction head box A. The suction heads A1 on the suction head box A located at the storage slots 324 are arranged in multiple rows, and each row of suction heads A1 is distributed in an arc shape concentric with the carrier plate 300. The first placement position, the second placement position and the liquid adding device are distributed on the carrier plate 300 along the circumference of the carrier plate 300.

[0127] During operation, the pipetting mechanism 500 can put the pipette tip A1 back into the pipette tip box A for temporary storage. Figure 8 As shown, a temporary pipette tip storage rack 216 may be provided at the top center of the reaction turntable 200 , and the temporary pipette tip storage rack 216 includes a circle of temporary pipette tip storage slots.

[0128] As attached Fig. 9 As shown, in order to facilitate power supply and communication, a mounting frame is provided on the carrier plate, and a second slip ring 325 concentric with the carrier plate is provided on the mounting frame.

[0129] As attached Fig.16 , Attachment Fig.17 As shown, the loading mechanism 400 includes a storage bin 401 and a pushing mechanism 402. The storage bin 401 is provided with a discharge port 403 at the bottom and highly matched with the radial slot 210 on the reaction turntable 200 on the side facing the reaction turntable 200. The distance between the discharge port 403 and the entrance of the radial slot directly facing it is less than the length of the reagent strip. The pushing member 404 of the pushing mechanism 402 can extend into the storage bin 401 and apply a thrust to the reagent strip B at the bottom of the storage bin 401 so as to push the reagent strip B at the bottom through the discharge port 403 to the radial slot 210 directly facing the discharge port 403.

[0130] As attached Fig.16 , Attachment Fig.17 As shown, the storage bin 401 has a rectangular storage cavity 405, and the reagent strips B are stored in the storage cavity 405 in a stacked manner. At the same time, the height of the discharge port 403 is slightly greater than the height of one of the reagent strips B, thereby ensuring that only the bottommost reagent strip B can be pushed out of the storage bin 401 at a time. The pushing member 404 of the pushing mechanism 402 can be a plate member with a thickness slightly smaller than the height of the discharge port 403. The plate member can extend into the storage bin 401 from the side of the storage bin 401 opposite to the discharge port, and when the pushing member pushes the reagent strip out, the pushing member can prevent the upper reagent strip from falling.

[0131] Preferably, in order to avoid the pushing member 404 from occupying too much horizontal space, the pushing member 404 is a vertical rod, and a through hole is provided at the bottom of the storage bin 401 to drive the vertical rod to move. The vertical rod is connected to the pushing drive mechanism 406, which can be a cylinder, a hydraulic cylinder, an electric cylinder or other feasible devices. Preferably, the pushing drive mechanism 406 is a structure composed of a motor and a lead screw, and the nut of the lead screw is connected to the pushing member.

[0132] As attached Fig.18 , Attachment Fig.19 As shown, in order to prevent the reagent strip B above from falling when the vertical rod pushes out the bottom reagent strip B, the loading mechanism 400 also includes a clamping mechanism 407, and the clamping mechanism 407 includes a roughly C-shaped rotating frame 408, and the rotating frame 408 can be rotatably arranged on the upper plate 110 around a rotating shaft 409 extending in the vertical direction. The rotating frame 408 includes a clamping portion 410 and a supporting portion 411 located at both ends thereof, and the height of the clamping portion 410 is equivalent to the height of the second lowest reagent strip B in the storage bin 401, and the side of the storage bin 401 is provided with a clamping operation hole 412 corresponding to the clamping portion 410. When the vertical rod moves away from the reaction turntable 200 until it contacts the abutment portion 411, the vertical rod can drive the rotating frame 408 to rotate around the rotating shaft 409, so that the pressing portion 410 is separated from the penultimate reagent strip B, so that the penultimate reagent strip B can fall to the bottom. When the vertical rod moves toward the reagent strip B and separates from the abutment portion 411, the rotating frame 408 is rotated and reset by the pressing spring 413 connected thereto, and the pressing portion 410 is pressed against the side of the penultimate reagent strip B through the pressing operation hole 412 to limit the reagent strip B from falling. Further, the vertical rod is provided with a roller matching the abutment portion 411.

[0133] As attached Fig.17As shown, in order to prevent the reagent strip B from tilting or falling to the bottom due to insufficient compression, which may interfere with the vertical rod that moves back to reset, the vertical rod is connected to a lifting cylinder 414 that drives it to move up and down. Of course, the lifting cylinder 414 can also use other devices that can achieve the same function, such as an electric cylinder, etc. The lifting cylinder is connected to the push drive mechanism 406. When the reagent strip B needs to be pushed out, the vertical rod rises to a high position, at which time the top of the vertical rod is higher than the bottom of the reagent strip at the bottom and not higher than the top of the reagent strip at the bottom. When the vertical rod needs to return to reset, the vertical rod drops to a low position, at which time the top of the vertical rod is lower than the reagent strip B at the bottom.

[0134] As attached Fig. 20 As shown, the injection assembly 501 includes an outer tube 502, which extends in the vertical direction and is arranged on a lifting frame 503. The lifting frame 503 is a feasible shape such as a C-shape or an I-shape, and includes a lower horizontal plate 504. The outer tube 502 is fixed below the lower horizontal plate 504. The upper horizontal plate 505 of the lifting frame 503 is connected to a lifting actuator 506 that drives it to rise and fall. The lifting actuator 506 can be a feasible device such as a cylinder, a hydraulic cylinder, an electric cylinder, and preferably a linear motor. The lifting actuator 506 is arranged on the side of a moving block 507 and is located above the lower horizontal plate. The bottom of the moving block 507 is sealed with an injection tube 508, and the injection tube 508 is inserted into the outer tube 502 and the two can move relative to each other. The lower end of the injection tube 508 is a cone 509, and a sealing ring is installed at the cone 509. The moving block 507 is provided with an injection cavity 510 which is connected with the liquid suction cavity of the injection tube 508 . The injection cavity 510 extends along the axial direction of the injection tube 508 and is provided with a piston rod 511 . The upper end of the piston rod 511 is connected to the upper horizontal plate 505 .

[0135] As attached Fig. 20 As shown, in order to ensure the sealing and stability of the piston rod 511, a guide sleeve 512 is provided at the upper end of the injection cavity 510, and the piston rod 511 is inserted into the guide sleeve 512. A sealing member 513 is also provided in the injection cavity 510 and is located below the guide sleeve 512. The sealing member 513 is sealingly sleeved on the outer periphery of the piston rod 511. An auxiliary roller 514 is also provided on the moving block 507, and the auxiliary roller 514 is attached to the inner side wall of the vertical plate of the lifting frame 503.

[0136] As attached Fig.21As shown, the loading drive mechanism 515 includes a lifting drive assembly 516 and a horizontal drive assembly 517. The lifting drive assembly 516 can be a linear module or a linear motor arranged in a vertical direction. Preferably, the moving block 507 can be slidably arranged on a longitudinal rail 518, and the lifting drive assembly 516 includes a belt 519 connected to the moving block 507, and the belt 519 is sleeved on an upper roller 520 and a lower roller 521, and one of the upper roller 520 and the lower roller 521 is connected to a lifting drive motor 522 that drives its rotation, and the lifting drive motor 522, the upper roller 520, the lower roller 521 and the longitudinal rail are arranged on a translation bracket 523. The translation bracket 523 is movably arranged on a horizontal slide rail 525 arranged on the top of a gantry 524 and connected to a horizontal drive assembly arranged on the gantry, and the gantry 524 is arranged across the top of the carrier plate 300. The horizontal drive assembly 517 can also be a linear module or a linear motor horizontally arranged on the gantry 524. In this embodiment, the structure of the horizontal drive assembly 517 is the same as that of the lifting drive assembly 516, which will not be described in detail. In addition, the sample loading drive mechanism 515 enables the injection tube 508 to move along the radial direction of the reaction turntable 200.

[0137] Since the sample needs to be added to the reaction cup B1 in the reaction chamber 204 through the liquid transfer mechanism 500, it is impossible to directly set the syringe of the liquid adding device directly above the reaction chamber 204 to add liquid. Therefore, when liquid needs to be added, the reagent strip B on the reaction turntable 200 should be pulled outwards for a certain distance so that the reaction cup B1 of the reagent strip B corresponds to the position of the avoidance hole. Therefore, in addition to being used to pull the reagent strip B from the reaction turntable 200 to the detection mechanism 700 for reading and unloading, the push-pull mechanism 600 can also pull the reagent strip B outwards for a certain distance from the radial slot 210 where it is located and push the reagent strip B to the inner end of the radial slot 210.

[0138] As attached Fig. 22 , Attachment Fig.23 As shown, the push-pull mechanism 600 includes a push-pull block 601, which is hinged on a translation block 602. The top of the push-pull block 601 is provided with a convex portion 603 located above the translation block. The convex portion 603 has an inclined surface 604 facing the reaction turntable 200, and the inclined surface 604 matches the lower vertex position of the outer end of the reagent strip B on the reaction turntable 200. The translation block 602 is connected to a push-pull driving mechanism 605 that drives it to reciprocate radially along the reaction turntable 200. The push-pull driving mechanism can be a known feasible device, preferably a linear motor and is arranged at the bottom of the upper plate. The translation block 602 can be slidably arranged on a guide rail 608 arranged on the upper plate.

[0139] When the inner end of the reagent strip B is inserted into the inner end of the radial slot 210 and cannot be inserted further, in the process of the inclined surface 604 contacting the lower vertex and continuing to move toward the center of the reaction turntable 200, the push-pull block 601 rotates relative to the translation block 602 to allow the protrusion 603 to rotate to the bottom of the reagent strip B.

[0140] As attached Fig.23 As shown, the push-pull block 601 is connected to the energy storage member 606, and the energy storage member 606 deforms and stores energy when the protrusion contacts and flips with the lower top corner of the reagent strip. When the protrusion moves to the push-pull groove B2 at the outer end of the reagent strip, the energy storage member causes the push-pull block to rotate in the opposite direction and reset, so that part of the protrusion is embedded in the push-pull groove B2. At the same time, the energy storage member 606 causes the push-pull block 601 to abut against the translation block 602 to limit the side 607 of the push-pull block 601 facing away from the reaction turntable 200 and extend in the vertical direction, so that when the push-pull block moves away from the center of the reaction turntable, the side 607 can push the reagent strip to move outward. The energy storage member 606 may be a torsion spring or a spring, etc., and when the reagent strip B is not completely inserted into the inner end of the radial slot 210, during the process of the protrusion 603 moving toward the center of the reaction turntable 200, the energy storage member 606 enables the protrusion to generate a thrust on the reagent strip B so that the reagent strip B is inserted into the inner end of the radial slot 210, so that the reaction cup B1 of the reagent strip B can be moved to the reaction chamber 204 again for reaction after the first solution and the second solution are added at the liquid adding device 301.

[0141] As attached Fig.23 , Attachment Fig.24 As shown, the detection mechanism 700 includes a stand 701 erected on the upper plate 110, and the bottom of the middle cross plate 702 of the stand 701 is provided with a receiving groove 703 corresponding to the height of the radial slot 210 and matching the position of the push-pull block, and the bottom of the middle cross plate 702 is provided with two receiving plates 704, and the two receiving plates 704 extend from both sides of the receiving groove 703 into the receiving groove 703 and keep a spacing between the two receiving plates 704 for the push-pull block 601 and the translation block to move through, and the two receiving plates 704 are bent downward toward one end of the reaction turntable to facilitate the reagent strip to enter the receiving groove. The middle cross plate 702 is provided with a reading hole 705 located above the receiving groove 703, and the middle cross plate 702 is provided with a quantum dot detector 706 located at the reading hole, and the quantum dot detector 706 is, for example, a known fluorescence detector.

[0142] In order to improve efficiency, there are two push-pull mechanisms and two detection mechanisms 700 , and each push-pull mechanism cooperates with one detection mechanism 700 .

[0143] After the reading is completed, the push-pull mechanism 600 will continue to move the reagent strip B away from the reaction turntable 200 so that the reagent strip B is pulled out from the receiving slot 703 of the detection mechanism 700. At this time, in order to facilitate the collection of waste, as shown in the attached Fig.24 As shown, a waste discharge chute C is also provided below the detection mechanism 700, and the waste discharge chute C is used to receive the reagent strip B extracted from the detection mechanism. The waste discharge chute C is provided from the top of the upper plate 110 to the bottom, and the lower end of the waste discharge chute is connected to the storage box D located below the reaction turntable 200, and the storage box D is limited at the limiting groove of the lower plate 120, and can be removed from the limiting groove. A material discharge hole (not shown in the figure) corresponding to the central hole of the storage box D and the inner ring is provided on the upper plate, and the material discharge hole is used for the pipetting mechanism 500 to put the used pipette tip A1 into the storage box D.

[0144] When using the automated testing device, first stack the prepared reagent strips in the storage bin; place at least one syringe 307 on the mounting block after sucking the first solution, and place at least one syringe 307 on the mounting block after sucking the second solution; and place the tip box A containing a group of tips A1 on the second placement position. Then, place the test tube containing the sample on the first placement position; specifically, insert one or more test tubes 910 containing the sample on the test tube rack 900, and then place the test tube rack 900 on the first placement position.

[0145] Then the S3 and S4 steps can be automatically performed.

[0146] The S3 includes the following process:

[0147] The reaction turntable 200 rotates until a radial slot 210 is opposite to the discharge port 403 of the loading mechanism 400, and the pushing mechanism 402 of the loading mechanism 400 is started to push the reagent strip B at the bottom of the storage bin 401 into the radial slot 210. At this time, the reaction cup B1 of the reagent strip B is located in the reaction bin 204 of the radial slot 210 and corresponds to the liquid adding hole 206.

[0148] The carrier plate 300 rotates to rotate the tip box A on it to the moving path of the syringe 508. The loading drive mechanism 515 drives the syringe 508 to move to just above a tip A1 and then moves the syringe 508 downward to be inserted into the tip A1. Then, the loading drive mechanism 515 drives the syringe 508 upward to remove the tip A1.

[0149] The reaction turntable 200 rotates to rotate the reagent strip B on it to a position corresponding to the pressure head 801. At the same time, the carrier plate 300 rotates to position a syringe 307 containing a first solution directly below the pressure head 801. At this time, the reagent strip B is opposite to the push-pull block of the push-pull mechanism. Then, the push-pull mechanism pulls the reagent strip B outward for a certain distance to make the reaction cup B1 on the reagent strip B directly face the syringe 307 below the pressure head 801. Then, the pressure head 801 moves downward to drive the piston core rod of the syringe 307 to move downward, thereby dripping a certain amount of the first solution into the reaction cup B1 on the reagent strip B. Then, the pressure head 801 is lifted, and the carrier plate 300 rotates to rotate a syringe 307 containing a second solution directly below the pressure head 801. Then, the pressure head 801 moves downward again to add the second solution into the reaction cup B1 of the reagent strip B. Next, the push-pull mechanism pushes the reagent strip B into the inner end of the radial slot 210 again. At this time, the push-pull block 601 is located in the push-pull through slot B2 at the outer end of the reagent strip B and does not affect the rotation of the reagent strip B. Moreover, each time the reaction turntable 200 rotates to a position, a radial slot 210 thereon is opposite to the discharge port 403 of the loading mechanism 400, so that the loading mechanism 400 can load the material.

[0150] The pipetting mechanism 500 draws the sample from the test tube in the first placement position and adds it to the reaction cup B1 of the reagent strip B. Specifically, the carrier plate 300 rotates until a test tube 910 containing a sample thereon is on the moving path of the injection tube 508, and then the pipetting mechanism 500 drives the injection tube 508 to move to just above the test tube 910 and then moves down to draw the sample in the test tube 910. At the same time, the reaction turntable 200 rotates to make the reagent strip B rotate to the moving path of the injection tube 508, and the pipetting mechanism 500 drives the injection tube 508 to move and adds the sample drawn by it to the reaction hole of the reagent strip B for reaction.

[0151] After the sample, the first solution and the second solution on a reagent strip B are dripped, the pipette mechanism 500 places the pipette tip A1 on the pipette tip temporary storage rack for temporary storage. At the same time, the pipette mechanism 500 can take another pipette tip A1 to add the sample of another reagent strip B. Of course, if only one sample needs to be tested, the pipette tip A1 does not need to be placed on the pipette tip temporary storage rack. Correspondingly, in the subsequent S4, there is no need to take the pipette tip from the pipette tip temporary storage rack.

[0152] The S4 includes the following process:

[0153] After a period of reaction, the sample, the first solution and the second solution on the reagent strip complete the reaction. After the pipetting mechanism 500 retrieves the pipette tip A1 corresponding to the reagent strip B from the pipette tip temporary storage rack, the reaction turntable 200 rotates to rotate the reaction cup B1 of the reagent strip B that has completed the reaction to the moving path of the injection tube 508. The pipetting mechanism 500 drives the injection tube 508 to the reaction cup B1 of the reagent strip B to absorb a certain amount of reaction liquid and drip it into the sample addition hole of the reagent strip B. Subsequently, the pipetting mechanism 500 moves the injection tube 508 to a position corresponding to the discharge hole, and then the lifting actuator 506 drives the outer tube 502 to move downward to push the pipette tip A1 at the lower end of the injection tube 508 downward away from the injection tube 508, so that the pipette tip A1 falls into the storage box D.

[0154] After chromatography for a period of time, the reaction turntable 200 rotates to rotate the reagent strip B to a position matching a push-pull mechanism 600, and the push-pull mechanism 600 draws the reagent strip B from the reaction turntable 200 to the detection mechanism 700 for reading.

[0155] After the detection mechanism 700 reads the value, the push-pull mechanism 600 extracts the reagent strip B from the detection mechanism 700. The reagent strip B extracted from the detection mechanism 700 separates from the push-pull mechanism 600 and falls into the waste discharge chute, and then slides down into the storage box D.

[0156] There are many implementation methods of the present invention, and all technical solutions formed by equivalent transformation or equivalent transformation fall within the protection scope of the present invention.

Claims

1. A method for detecting hs-TnI for non-disease diagnosis purposes based on quantum dot microspheres, characterized in that: The steps include: S1, preparing a reagent strip, wherein the reagent strip comprises a sample pad, an NC detection membrane, and absorbent paper which are sequentially arranged on a substrate, wherein the T line of the NC detection membrane is fixed with streptavidin, and the C line is fixed with mouse anti-DNP monoclonal antibody; S2, preparing a first solution and a second solution, wherein the first solution comprises quantum dot microspheres coupled to troponin antibody 1 and quantum dot microspheres coupled to DNP-BSA, and the second solution comprises troponin antibody 2; S3, mixing a certain sample, the first solution and the second solution and reacting them for a certain period of time to obtain a reaction solution; S4, taking a certain amount of reaction solution and dropping it into the sample adding hole of the reagent strip for chromatography for a certain period of time, and then detecting the T line and C line of the reagent strip by a quantum dot detector to obtain the detection result; S3 and S4 are implemented by an automated testing device, which includes a base, on which are provided: A reaction turntable, which is rotatably disposed on the base relative to the base and connected to a first rotating mechanism that drives the reaction turntable to rotate, and is provided with a radial slot for inserting a reagent strip radially therefrom; A carrier plate, which is concentric with the reaction turntable and is arranged above the reaction turntable, and the carrier plate is provided with a liquid adding device for adding a first solution and a second solution to the reagent strip on the reaction turntable; the carrier plate is provided with a first placement position for placing a test tube containing a sample and a second placement position for placing a tip box containing a tip; A loading mechanism is arranged at the periphery of the reaction turntable and is used to insert the reagent strips one by one into the radial slots of the reaction turntable; the loading mechanism includes a storage bin and a pushing mechanism, the storage bin is provided with a discharge port at the bottom and highly matched with the radial slots on the reaction turntable on one side of the storage bin facing the reaction turntable, and a pushing member of the pushing mechanism can extend into the storage bin and apply a thrust to the reagent strip at the bottom in the storage bin so as to push the reagent strip at the bottom through the discharge port to the radial slot directly opposite to the discharge port; The liquid transfer mechanism comprises an injection assembly and a sample adding drive mechanism for driving the injection assembly to move horizontally and vertically, wherein the liquid transfer mechanism is used to draw a sample from a test tube containing a sample at a first placement position and add the sample to a reaction cup of a reagent strip on a reaction turntable, and to draw a reaction liquid in the reaction cup and add the reaction liquid to a sample adding hole of the reagent strip; A push-pull mechanism, arranged at the periphery of the reaction turntable, for enabling the reagent strip on the reaction turntable to be transferred from the radial slot where the reagent strip is located to the detection mechanism and to be withdrawn from the detection mechanism; A detection mechanism, arranged beside the push-pull mechanism, for reading the reagent strip inserted thereon; The push-pull mechanism comprises a push-pull block, the push-pull block is hinged on a translation block, a convex portion is arranged on the top of the push-pull block, the convex portion has an inclined surface facing the reaction turntable, the inclined surface matches the lower vertex position of the outer end of the reagent strip on the reaction turntable so that after the inclined surface moves and contacts the lower vertex, the convex portion rotates to the bottom of the reagent strip; the push-pull block is connected to an energy storage member, the energy storage member makes the push-pull block abut against the translation block to limit the position so that the push-pull block extends in the vertical direction against the side of the reaction turntable, and the translation block is connected to a push-pull drive mechanism that drives it to reciprocate radially along the reaction turntable; The liquid adding device comprises a mounting block, a group of limiting holes for mounting a syringe filled with liquid medicine are arranged at the mounting block, avoidance holes corresponding to the limiting holes are arranged on the carrying plate, the limiting holes are arranged in an arc shape, and the arc shape of the limiting holes is concentric with the carrying plate; the carrying plate is arranged on the base so as to be rotatable relative to the base and is connected to a second rotating mechanism driving its rotation; The liquid adding device, the first placement position and the second placement position are distributed on the carrier plate along the circumference of the carrier plate; a pressing device for driving the piston core rod of the syringe at the liquid adding device to move downward is arranged on the outer side of the carrier plate.

2. The method for detecting hs-TnI for non-disease diagnosis purposes based on quantum dot microspheres according to claim 1, characterized in that: The preparation process of the first solution is as follows: S21, preparing a first quantum dot microsphere solution including quantum dot microspheres coupled to troponin antibody 1; S22, preparing a second quantum dot microsphere solution including quantum dot microspheres coupled with DNP-BSA; S23, diluting the first quantum dot microsphere solution and the second quantum dot microsphere solution respectively with microsphere buffer and mixing them to obtain a first solution.

3. The non-disease diagnosis purpose detection method of hs-TnI based on quantum dot microspheres according to claim 2, characterized in that: The sealing agent used in the preparation process of the first quantum dot microsphere solution and the second quantum dot microsphere solution is CE510 sealing agent.

4. The method for detecting hs-TnI for non-disease diagnosis purposes based on quantum dot microspheres according to claim 2, characterized in that: The microsphere buffer includes sucrose and Tween 20.

5. The method for detecting hs-TnI for non-disease diagnosis purposes based on quantum dot microspheres according to claim 1, characterized in that: The particle size of the quantum dot microspheres is greater than or equal to 10 nanometers and less than 100 nanometers.

6. The method for detecting hs-TnI for non-disease diagnosis purposes based on quantum dot microspheres according to claim 1, characterized in that: The OD of the first solution 340 Between 0.10-0.

20.

7. The method for detecting hs-TnI for non-disease diagnosis purposes based on quantum dot microspheres according to claim 1, characterized in that: The troponin antibody 2 is conjugated to biotin.

8. The method for detecting hs-TnI for non-disease diagnosis purposes based on quantum dot microspheres according to claim 1, characterized in that: In S3, the sample, the first solution and the second solution are allowed to react at a constant temperature for 3-5 minutes.

9. The method for detecting hs-TnI for non-disease diagnosis purposes based on quantum dot microspheres according to claim 1, characterized in that: In the S4, the chromatography time is 4-6 minutes.

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