D-dimer detection kit and detection method thereof

Abstract

The present application relates to the technical field of medical detection, in particular to a D-dimer detection kit and a detection method thereof.The present application provides a D-dimer detection kit, which is composed of reagent R1 and reagent R2.The reagent R1 is mainly composed of a buffer, a preservative, an inorganic salt, a surfactant and a protein stabilizer;the reagent R2 is mainly composed of a buffer, a protein protective agent, sucrose and a preservative.The experiment shows that the kit is easy to operate and has a short analysis time, and the kit also has high analysis sensitivity, optimal precision, good test accuracy and a wide linear range, and can be combined with an automatic analyzer, which is conducive to its wide use in clinical practice.

Description

Technical Field

[0001] This invention relates to the field of medical testing technology, specifically to a D-dimer detection kit and its detection method. Background Technology

[0002] D-dimer (DD) is a specific fibrin degradation product produced by plasmin under the action of fibrinolytic enzymes. During blood clotting, fibrinogen is hydrolyzed into soluble fibrin monomers, which spontaneously polymerize into network polymers. Under the action of thrombin and factor XIIIa, these monomers covalently cross-link to form cross-linked fibrin. Simultaneously, the cross-linked fibrin in the body is degraded by plasmin into fibrin degradation products of various molecular weights, including its specific degradation product, D-dimer.

[0003] Activation of the coagulation system leads to the generation of thrombin. After the central domain (E region) of fibrinogen is bound by thrombin, fibrin peptide A (FPA) and fibrin peptide B (FPB) are released, simultaneously exposing complementary binding sites to facilitate their spontaneous polymerization. Fibrin units polymerize side-by-side and end-to-end to form polymer chains, generating fibrin monomers (FM) and polymers. In Factor XIIIa and Ca2+... 2+ Under the influence of [the enzyme / organism], glutamine at position 398 or 399 of the γ chain on a fibrin monomer cross-links with lysine at position 406 of the γ chain on another fibrin monomer to form a γ'-γ' bond, generating cross-linked fibrin. Plasmin cleaves the complementary structure of the DE region, degrading fibrinogen (Fg) and fibrin monomer (FM) to generate fibrinogen degradation products (Fg DPs); simultaneously, it de-cross-linked fibrin generates various cross-linked fibrin degradation products (Fb DPs), which include D-dimers and other fragments.

[0004] Elevated D-dimer levels can detect the progression of various diseases, such as deep vein thrombosis, disseminated intravascular coagulation, myocardial infarction, severe hepatitis, and pulmonary embolism. It also has a certain monitoring role in potential complications in preeclampsia and high-risk pregnancies. Furthermore, changes in D-dimer levels can serve as an indicator for monitoring thrombolytic therapy and guiding the dosage of thrombolytic drugs. Therefore, detecting D-dimer levels in the blood has significant clinical value in the early diagnosis, disease progression monitoring, and treatment monitoring of thrombolytic drugs in thrombotic diseases.

[0005] Currently, there are three main methods for D-dimer testing: whole blood agglutination assay, enzyme-linked immunosorbent assay (ELISA) & enzyme-linked immunofluorescence assay (ELFA), and latex immunoturbidimetry. Among them, latex immunoturbidimetry is widely used in clinical practice due to its advantages of high sensitivity, rapid detection, and accuracy. The detection of D-dimer using latex immunoturbidimetry is based on the binding of the analyte D-dimer to antibodies coated on the surface of latex particles, causing an increase in turbidity, thereby achieving the detection of D-dimer. However, current kits for detecting D-dimer suffer from low precision, narrow linear range, weak anti-interference ability, and poor stability, which limits their application in clinical testing. Summary of the Invention

[0006] In view of this, the technical problem to be solved by the present invention is to provide a D-dimer detection kit and a detection method thereof.

[0007] This invention provides a kit for D-dimer detection, comprising reagent R1 and reagent R2.

[0008] The reagent R1 includes: buffer solution, inorganic salt, coagulant, surfactant, preservative and protectant; the surfactant includes at least one of: organosilicon surfactant, fluorosurfactant and / or nonionic surfactant;

[0009] Furthermore, the silicone surfactant includes, but is not limited to, Silwet L-7600; the nonionic surfactant includes, but is not limited to, Triton X-100.

[0010] The reagent R2 comprises: buffer, protectant, stabilizer and latex microspheres coated with D-Dimer antibody;

[0011] The latex microspheres are made of polystyrene latex microspheres with a particle size of 300 nm.

[0012] The present invention optimizes the particle size of the latex microspheres. Experimental results show that when the particle size of the carboxylated polystyrene latex microspheres of the present invention is selected as 300 nm, the monoclonal antibody affinity is good, and the reactivity and sensitivity are high.

[0013] Furthermore, in reagent R1,

[0014] The buffer solution is selected from at least one of MOPS buffer and / or PB buffer;

[0015] The inorganic salt is selected from at least one of NaCl and / or KCl;

[0016] The coagulant is selected from at least one of PEG6000 and / or PEG8000;

[0017] The surfactants include Silwet L-7600 and Triton X-100;

[0018] The preservative is ProClin 300;

[0019] The protective agent is selected from at least one of BSA, Casein and / or ADP;

[0020] In reagent R2,

[0021] The buffer solution is selected from at least one of MOPS buffer, TAPS buffer, PB buffer, glycine buffer and / or HEPES buffer;

[0022] The protective agent is selected from at least one of BSA, Casein and / or ADP;

[0023] The stabilizer is selected from at least one of sucrose, trehalose, glycerol and / or glucose.

[0024] Furthermore,

[0025] The reagent R1 includes: 50 mM PB buffer at pH 6.5, 9–15 g / L NaCl, 8–10 g / L PEG6000, 1.0–2.0 g / L Silwet L-7600, 1.0–5.0 g / L Triton X-100, 1.0–2.0 ml / L ProClin 300, and 2–5 g / L BSA.

[0026] The reagent R2 comprises: 20 mM MOPS buffer, 2–5 g / L BSA, 60–80 g / L glucose, and 1–3 mg / mL polystyrene latex microspheres coated with D-Dimer antibody; or

[0027] The reagent R2 comprises: 20 mM TAPS buffer, 2–5 g / L BSA, 60–80 g / L glucose, and 1–3 mg / mL polystyrene latex microspheres coated with D-Dimer antibody; or

[0028] The reagent R2 comprises: 20 mM HEPES buffer, 2–5 g / L BSA, 60–80 g / L glucose, and 1–3 mg / mL polystyrene latex microspheres coated with D-Dimer antibody.

[0029] Furthermore,

[0030] In some specific embodiments of the present invention, reagent R1 is 50 mM PB buffer at pH 6.5, 9 g / L NaCl, 8 g / L PEG6000, 1.0 g / L Silwet L-7600, 1.0 g / L Triton X-100, 1.0 mL / L ProClin 300, and 5 g / L BSA; reagent R2 is 20 mM MOPS buffer, 2 g / L BSA, 60 g / L glucose, and 1.25 mg / mL polystyrene latex microspheres coated with D-Dimer antibody; or

[0031] In other embodiments of the present invention, reagent R1 is 50 mM PB buffer at pH 6.5, 12 g / L NaCl, 9 g / L PEG6000, 1.0 g / L Silwet L-7600, 2.0 g / L Triton X-100, 1.0 mL / L ProClin 300, and 2 g / L BSA; reagent R2 comprises: 20 mM TAPS buffer, 2 g / L BSA, 60 g / L glucose, and 1.25 mg / mL polystyrene latex microspheres coated with D-Dimer antibody; or

[0032] In other embodiments of the present invention, reagent R1 is 50 mM PB buffer at pH 6.5, 15 g / L NaCl, 10 g / L PEG6000, 2.0 g / L Silwet L-7600, 5.0 g / L Triton X-100, 1.0 mL / L ProClin300, and 2 g / L BSA; reagent R2 comprises: 20 mM HEPES buffer, 5 g / L BSA, 80 g / L glucose, and 1.25 mg / mL polystyrene latex microspheres coated with D-Dimer antibody.

[0033] In the kit described in this invention, the surfactant in reagent R1 is a combination of the highly efficient organosilicone surfactant Silwet L-7600 and the nonionic surfactant 100 (polyethylene glycol octylphenyl ether) (Triton X-100). Organosilicone surfactants not only have higher surface activity (e.g., lower CMC and γCMC) than ordinary surfactants, but also possess many properties that ordinary surfactants do not have. The surfactant in reagent R1 acts as a dispersant, allowing for more complete exposure of the antigen site, thereby promoting the attraction and binding between the analyte D-dimer and the latex particles of the D-dimer antibody. This effectively solves the problem of poor repeatability at low values, improves the accuracy of reagent detection, and significantly enhances the reagent's anti-interference ability.

[0034] In this invention, the surfactant in R1 effectively solves the problem of reagent reproducibility. In some specific embodiments, the surfactant in reagent R1 is preferably a combination of the highly efficient organosilicon surfactant Silwet L-7600 and the conventional surfactant 100 (polyethylene glycol octylphenyl ether) (Triton X-100). On the one hand, the combination of the buffer solution of reagent R1 with the above-mentioned surfactant can effectively solve the problem of reagent reproducibility. In some specific experimental groups, the buffer solution of reagent R1 is preferably a 50mM PB buffer solution with pH 6.5. On the other hand, the addition of sodium chloride in the presence of surfactant and other components in R1 further enhances the reagent's anti-interference ability.

[0035] This invention's kit is based on immunoturbidimetric assay for D-dimer determination. The detection principle is as follows: reagent R2 combines with D-dimer in the sample, causing an agglutination reaction to form an antigen-antibody immune complex, producing turbidity. The level of turbidity is related to the concentration of D-dimer in the sample. The absorbance value at this point is measured, and the D-dimer content is calculated based on a calibration curve. Typically, the particle size of the polystyrene latex microspheres affects the reagent's sensitivity. The polystyrene latex microspheres have a particle size of 270 nm to 400 nm; in some specific embodiments, the polystyrene latex microspheres have a particle size of 300 nm.

[0036] This invention provides a method for detecting D-dimer, which uses the kit described in this invention to detect samples.

[0037] Furthermore, the detection includes the following steps: after the sample is mixed with reagent R1 and reagent R2 in sequence, the difference between the initial absorbance and the measured absorbance is obtained, and the concentration of the D-dimer is obtained according to the standard curve.

[0038] Furthermore,

[0039] The volume ratio of the sample, reagent R1, and reagent R2 is 150:50:6.

[0040] The initial absorbance was measured 5 minutes after reagent R2 was mixed;

[0041] The absorbance was measured 10 minutes after reagent R2 was mixed.

[0042] The sample was plasma, which had been treated with sodium citrate.

[0043] Specifically, reagent R1 is first mixed with the sodium citrate anticoagulated plasma sample to be tested for 5 minutes, and then reagent R2 is added to start the reaction. The volume ratio of reagent R1:reagent R2:sample is 150:50:6. After 5 minutes, the initial absorbance A1 is measured by comparing with the blank, and then the absorbance A2 is measured by comparing with the blank at 10 minutes. The difference between A2 and A1, ΔA, is calculated, and the content of D-dimer in the plasma sample to be tested is obtained according to the standard curve.

[0044] This invention provides a D-dimer detection kit, which consists of reagent R1 and reagent R2. Reagent R1 mainly consists of buffer solution, preservative, inorganic salt, surfactant, and protein stabilizer; reagent R2 (latex reagent) mainly consists of buffer solution, protein protectant, sucrose, and preservative. Experiments show that this kit is not only easy to operate and has a short analysis time, but also possesses high analytical sensitivity, excellent precision, good test accuracy, and a wide linear range. Furthermore, it can be integrated with automated analyzers, facilitating its widespread clinical application. Attached Figure Description

[0045] Figure 1 The correlation curve of experimental group 1 in the correlation experiment;

[0046] Figure 2 The linear range curve of experimental group 1 is shown in the linear experiment. Detailed Implementation

[0047] This invention provides a D-dimer detection kit and its detection method. Those skilled in the art can refer to the content of this document and appropriately modify the process parameters to achieve the same result. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The methods and applications of this invention have been described through preferred experimental groups. Those skilled in the art can clearly modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0048] The test materials used in this invention are all common commercial products and can be purchased on the market.

[0049] The invention will be further elaborated below with reference to the experimental group:

[0050] Experimental Group D-dimer Detection Kit and Detection Method

[0051] I. Preparation of Reagent R1: The kit of this invention is based on latex immunoturbidimetric assay for D-dimer determination and consists of reagent R1 and reagent R2. Reagent R1 contains a combination of surfactants Silwet L-7600 and Triton X-100. Reagent R2 is a polystyrene latex microsphere complex coated with D-dimer antibody.

[0052] Table 1. Components of Reagent R1

[0053]

[0054]

[0055] The preparation steps for reagent R1 are as follows (preparing 1L): According to the component concentrations of experimental groups 1 to 3 in Table 1, and according to the component content of reagent R1 mentioned above, prepare PB buffer using purified water, adjust the pH, and use it as R1 buffer; then add the corresponding amounts of sodium chloride, polyethylene glycol 6000, Silwet L-7600, Triton X-100, bovine serum albumin, and ProClin300, mix well, and adjust the pH with sodium hydroxide / hydrochloric acid to obtain reagent R1.

[0056] II. Preparation of reagent R2:

[0057] Table 2. Components of reagent R2

[0058] Components Experimental group 1 Experimental group 2 Experimental group 3 20mM buffer MOPS TAPS HEPES Bovine serum albumin (BSA, g / L) 2 2 5 Glucose (g / L) 60 60 80 Latex microspheres coated with DD antibody (g / L) 1.25 1.25 1.25

[0059] 1. Preparation of buffer solution for reagent R2: Prepare buffer solution using purified water according to the component concentrations of reagent R2 as shown in Table 2, adjust the pH, and then add the corresponding amounts of bovine serum albumin and glucose to prepare the buffer solution for reagent R2.

[0060] 2. Preparation of reagent R2:

[0061] Step 1: Add carboxylated polystyrene latex microspheres with a particle size of 300 nm to 10 mM MES buffer to make the concentration 8.3 mg / ml;

[0062] Step 2: Add 4 mg / ml EDC ((1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride)) to the above latex microspheres and stir at room temperature for 0.5 h;

[0063] Step 3: Dissolve and dilute the DD monoclonal antibody to 0.24 mg / ml with PB buffer at pH 8.0, and then add the antibody dilution solution to the activated polystyrene latex microspheres and stir at room temperature for 3 hours.

[0064] Step 4: Add commercial blocking agent 110010 to terminate the reaction, bringing the final concentration to 0.05%, and stir overnight at room temperature; centrifuge the resulting reaction solution, discard the supernatant, and obtain the precipitate;

[0065] Step 5: Disperse the precipitate with buffer solution of reagent R2 and sonicate for 3 minutes to obtain uniformly distributed R2 latex reagent;

[0066] Step 6: Place the above R2 in a 42℃ water bath for 16-18 hours to age it and obtain the final R2 reagent.

[0067] III. Preparation of reagent kits for control groups 1-3

[0068] The preparation of reagent R1 is the same as that of one of experimental groups 1 to 3.

[0069] Preparation of reagent R2: Except for the different particle size of polystyrene latex microspheres used in step 1 of the experimental group, the particle sizes of microspheres used in control groups 1 to 3 were 145 nm, 190 nm and 218 nm, respectively. All other steps were consistent with the experimental group.

[0070] IV. Preparation of reagent kits for control groups 4-9

[0071] The kit is used for the determination of D-dimer by latex immunoturbidimetric assay and includes reagents R1 and R2.

[0072] Table 3. Components of Reagent R1

[0073]

[0074] The preparation steps for reagent R1 are as follows (preparing 1L): Prepare a buffer solution using purified water according to the component concentrations of control groups 4-9 in Table 3 and the component content of reagent R1 mentioned above, and adjust the pH to obtain the R1 buffer solution; then add the corresponding amounts of sodium chloride, polyethylene glycol 6000, Silwet L-7600, Triton X-100, bovine serum albumin, and ProClin300, mix well, and adjust the pH with sodium hydroxide / hydrochloric acid to obtain reagent R1.

[0075] The preparation method of reagent R2 is the same as that of reagent R2 in the experimental group.

[0076] V. Results

[0077] The performance evaluation results of the reagent kits described in experimental groups 1 to 3 of this invention are basically consistent. Taking the reagent kits prepared in experimental groups 1 to 3 as examples, the correlation, linear range, precision, anti-interference, stability and other related performances are verified.

[0078] Test method: Endpoint method, incremental reaction. At 37℃, reagent R1 was first mixed with the DD plasma sample for 5 minutes, and then reagent R2 was added to start the reaction, with a volume ratio of reagent R1:reagent R2:sample = 150:50:6. The initial absorbance A1 was measured relative to the blank at 5 minutes, and then the absorbance A2 was measured relative to the blank at 10 minutes. The difference between A2 and A1, ΔA, was calculated.

[0079] 1. Determination of the standard curve

[0080] The ΔA values ​​of standard plasma samples with a standard concentration of DD ranging from 0 to 17.6 μg / mL were determined using reagent kits for experimental groups 1–3 and the control DD reagent kit (SEKISUID-dimer kit, catalog number: 346903). Partial concentration detection results are shown in Table 4. Standard curves for each reagent kit were fitted based on the concentrations of DD and the measured ΔA values. The standard curves for experimental groups 1–3 were fitted using the SPLINE function.

[0081] Table 4 shows the correspondence between the concentration of DD and the measured ΔA value. As can be seen from Table 4, under the same conditions, the absorbance of experimental groups 1-3 changed more significantly and the sensitivity was higher than that of control groups 1-3.

[0082] Table 4. ΔA value test results of standard plasma samples

[0083]

[0084] As can be seen from Table 4, compared with the experimental groups 1-3, the reactivity of control groups 1-3 was lower and the precision of the test samples was worse (Table 7). This indicates that the particle size of the carboxylated polystyrene latex microspheres of the present invention was appropriate, the monoclonal antibody used had good affinity, and the reactivity and sensitivity could reach the level of the control kit.

[0085] 2. Correlation Experiment

[0086] A commercially recognized D-dimer kit with excellent accuracy was used as a control example (SEKISUID-dimer kit, catalog number: 346903). The kits used in the experimental group were used as the experimental group for comparison. Forty samples were tested, and the results are shown in Table 5.

[0087] Using the test results of commercially available control kits as the independent variable on the x-axis and the test results of the kit of this invention as the dependent variable on the y-axis, linear regression curves were plotted. The regression equation for experimental group 1 was Y = 1.031X - 0.0524, with a linear correlation coefficient R = 0.9971; the regression equation for experimental group 2 was Y = 1.0251X + 0.0767, with a linear correlation coefficient R = 0.9990; and the regression equation for experimental group 3 was Y = 1.0263X + 0.0392, with a linear correlation coefficient R = 0.9970. The linear relationships were good, and the test results can be effectively used for clinical testing. The correlation curve for experimental group 1 is shown below. Figure 1 .

[0088] Table 5. Results of correlation test (unit: μg / mL)

[0089]

[0090]

[0091] 3. Precision testing

[0092] Two plasma samples with different DD concentrations were taken and tested 20 times consecutively. The coefficient of variation was calculated. The precision results of the experimental group are shown in Table 6, and the precision results of the control groups 1-9 are shown in Table 7. From the experimental results of experimental groups 1-3 and control groups 1-3 in Tables 6 and 7, it can be seen that compared with reagent 2 prepared by 145nm, 190nm and 218nm microspheres, the use of 300nm microspheres has higher sensitivity in the low value region and better repeatability of the test samples.

[0093] The experimental results of experimental groups 1-3 and control groups 4-7 in Tables 6 and 7 show that adding a common surfactant to reagent R1 can improve the precision of the reagent, while adding a silicone surfactant can significantly improve the precision. Furthermore, the addition of common surfactants and special silicone surfactants has a synergistic effect in improving precision. Also, the experimental results of experimental groups 1-3 and control groups 8-9 in Tables 6 and 7 show that the buffer type affects the precision of the reagent, with 50 mM PB being the preferred choice.

[0094] Table 6. Precision test results for experimental groups 1–3

[0095]

[0096] Table 7. Precision test results for comparison groups 1–9

[0097]

[0098] 4. Anti-interference detection

[0099] Serum from clinically healthy patients was divided into two portions. One portion was treated with the highest concentration of the interfering substance, while the other portion was treated with an equal volume of solvent. Samples with and without the interfering substance were subjected to three graded differential dilutions. Each sample was tested three times, and the deviation of the measurements was calculated. The anti-interference ability of the experimental group was evaluated, and the results are shown in Table 8. The anti-interference ability of control groups 4–7 was also evaluated, and the results are shown in Table 9.

[0100] Table 8. Anti-interference test results for experimental groups 1-3

[0101]

[0102] Table 9. Anti-interference test results for comparison groups 4–7

[0103]

[0104]

[0105] The results showed that the reagent kit of this invention exhibited strong anti-interference performance, with resistance to interference from bilirubin, hemoglobin, fat emulsion, vitamin C, and heparin sodium at three concentration gradients all within ±5%. In contrast, control groups 4-7 showed poor anti-interference performance, with resistance to hemoglobin and chyle interference exceeding 10%. These experimental results indicate that the addition of surfactants Silwet L-7600 and Triton X-100 significantly enhances the reagent's anti-interference ability. Furthermore, the reagent demonstrated optimal anti-interference performance with the addition of sodium chloride. Therefore, the combination of surfactants Silwet L-7600 and Triton X-100, and their combined use with sodium chloride, can more significantly improve the reagent's anti-interference ability.

[0106] 5. Linear Experiment

[0107] Seven dilutions were prepared by mixing high- and low-value samples (in equal proportions), and each sample was measured twice and the mean was taken. Normal samples were mixed to obtain a low-value sample of 0.32 μg / mL. A high-value DD sample of 17 μg / mL was added to the mixed normal samples as the linear high value. The high- and low-value samples were then mixed proportionally to create seven equally diluted samples. The linearity results are shown in Table 10. Using theoretical DD concentration as the independent variable X on the x-axis and actual measured value as the dependent variable Y on the y-axis, a linear regression equation was derived, and the correlation coefficient R was calculated. The results showed that the linear regression equation for experimental group 1 was Y = 0.9955X - 0.0419, with a correlation coefficient R = 0.9998; the linear regression equation for experimental group 2 was Y = 1.0093X - 0.01, with a correlation coefficient R = 0.9999; and the linear regression equation for experimental group 3 was Y = 1.0109X - 0.0207, with a correlation coefficient R = 0.9998. This indicates that the present invention exhibits good correlation within the linear range of 0.3 μg / mL to 17 μg / mL. The linear range curve for experimental group 1 is shown below. Figure 2 .

[0108] Table 10. Results of Linear Range Analysis

[0109]

[0110] 6. Stability Test

[0111] The reagent kit of the present invention was subjected to open-bottle stability and long-term stability tests. The reagent kit of the present invention was calibrated on the testing instrument, and stored at 2-8℃ for 30 days after opening. After 30 days, open-bottle stability tests were performed on plasma samples with concentrations of 4.0 μg / mL and 8.0 μg / mL, respectively. The deviation value of the test results after 30 days of opening was calculated, and the results are shown in Table 11.

[0112] Table 11. Results of bottle-opening stability test for experimental groups 1-3

[0113]

[0114] The reagent kit of this invention was calibrated on the testing instrument and stored in a sealed container at 2–8°C for 18 months. Long-term stability tests were conducted on plasma samples with concentrations of 4.0 μg / mL and 8.0 μg / mL at 3, 6, 9, 12 and 18 months, respectively. The deviation value of the test results after 18 months of opening was calculated. The results are shown in Table 12.

[0115] Table 12. Long-term stability test results for experimental groups 1–3 (unit: μg / mL)

[0116]

[0117] The results in Tables 11 and 12 show that, after 30 days of opening and 18 months of sealed storage at 2–8°C, the test deviations for plasma samples with concentrations of 4.0 μg / mL and 8.0 μg / mL using the reagent kit of this invention are both within 3%, indicating good stability. The reagent R2 prepared in this invention exhibits good stability.

[0118] The above are merely preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A kit for D-dimer detection, characterized in that, Including reagent R1 and reagent R2, The reagent R1 includes: PB buffer, NaCl, coagulant, surfactant, preservative and protective agent; the surfactant includes Silwet L-7600 and Triton X-100; The reagent R2 comprises: buffer, protectant, stabilizer and latex microspheres coated with D-Dimer antibody; The latex microspheres are made of polystyrene latex microspheres with a particle size of 300 nm.

2. The reagent kit according to claim 1, characterized in that, In reagent R1, The coagulant is selected from at least one of PEG6000 and / or PEG8000; The preservative is ProClin 300; The protective agent is selected from at least one of BSA, Casein and / or ADP.

3. The reagent kit according to claim 1, characterized in that, In reagent R2, The buffer solution is selected from at least one of MOPS buffer, TAPS buffer, PB buffer, glycine buffer and / or HEPES buffer; The protective agent is selected from at least one of BSA, Casein and / or ADP; The stabilizer is selected from at least one of sucrose, trehalose, glycerol and / or glucose.

4. The kit according to claim 1 or 2, characterized in that, The reagent R1 comprises: 50 mM PB buffer at pH 6.5, 9–15 g / L NaCl, 8–10 g / L PEG6000, 1.0–2.0 g / L Silwet L-7600, 1.0–5.0 g / L Triton X-100, 1.0–2.0 ml / L ProClin 300, and 2–5 g / L LBSA.

5. The kit according to claim 1 or 3, characterized in that, The reagent R2 comprises: 20 mM MOPS buffer, 2-5 g / L BSA, 60-80 g / L glucose, and 1-3 mg / mL polystyrene latex microspheres coated with D-Dimer antibody; or The reagent R2 comprises: 20 mM TAPS buffer, 2-5 g / L BSA, 60-80 g / L glucose, and 1-3 mg / mL polystyrene latex microspheres coated with D-Dimer antibody; or The reagent R2 comprises: 20 mM HEPES buffer, 2-5 g / L BSA, 60-80 g / L glucose, and 1-3 mg / mL polystyrene latex microspheres coated with D-Dimer antibody.

6. A method for detecting D-dimer for non-diagnostic purposes, characterized in that, The sample is tested using the kit described in any one of claims 1 to 5.

7. The detection method according to claim 6, characterized in that, The detection includes the following steps: after the sample is mixed with reagent R1 and reagent R2 in sequence, the difference between the initial absorbance and the measured absorbance is obtained, and the concentration of the D-dimer is obtained according to the standard curve.

8. The detection method according to claim 6 or 7, characterized in that, The volume ratio of the sample, reagent R1, and reagent R2 is 150:50:

6.

9. The detection method according to claim 6 or 7, characterized in that, The initial absorbance was measured 5 minutes after reagent R2 was mixed; The absorbance was measured 10 minutes after reagent R2 was mixed.

10. The detection method according to any one of claims 6 to 9, characterized in that, The sample was plasma, which had been treated with sodium citrate.

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