A detection kit for matrix metalloproteinase-3 based on latex immunoturbidimetry

Abstract

The invention discloses a matrix metalloproteinase-3 detection kit based on latex immunoturbidimetry, and belongs to the field of biotechnology. The invention adds a composite sensitizer composed of dextran, polyethylene glycol 4000 and tributylmethylammonium chloride to reagent R1, and adds a composite stabilizer composed of lithium chloride, hydroxylamine hydrochloride and a protective agent to reagent R2, and improves the detection sensitivity and the dispersion stability of latex particles through the different action mechanisms of each component, thereby providing a matrix metalloproteinase-3 detection kit based on latex immunoturbidimetry with high sensitivity and high stability.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a matrix metalloproteinase-3 detection kit based on latex immunoturbidimetry. Background Art

[0002] Rheumatoid arthritis (RA) is a common autoimmune disease characterized by chronic inflammatory lesions of the synovial membrane. Its main pathological features are persistent synovial inflammation and cell infiltration, which eventually affect cartilage and bone, leading to joint structural destruction, deformity and dysfunction, resulting in varying degrees of disability. The pathogenesis of RA is still not fully understood. Matrix metalloproteinase-3 (MMP-3) is an enzyme that decomposes the synovium. Studies have found that elevated levels of MMP-3 destroy cartilage. MMP-3 is expressed at high levels in the serum of RA patients, and may play an important role in the progressive joint destruction of RA.

[0003] The level of MMP-3 in the joint membrane is related to the level of MMP-3 in the serum, so the level of MMP-3 in the serum can be used to check the condition of the patient's joints. Clinical or laboratory diagnostic methods include: radioimmunoassay, enzyme-linked immunosorbent assay and latex immunoturbidimetry. Among them, the principle of latex immunoturbidimetry is that MMP-3 in serum and its corresponding antibody-sensitized latex particles meet in the solution, agglutination reaction occurs, and a certain turbidity is produced. The level of turbidity is proportional to the concentration of the antigen in the presence of a certain amount of antibody. At a fixed wavelength, by comparing with the same treated calibrant, the concentration of MMP-3 in the sample can be detected. Matrix metalloproteinase-3 latex immunoturbidimetry kits are commonly used in the market to measure MMP-3, and stability and sensitivity are the main indicators of concern.

[0004] The patent (CN106950382A) discloses a matrix metalloproteinase-3 assay reagent, including a pretreatment reagent, an antibody latex reagent, and a calibration reagent; the pretreatment reagent includes a buffer, a preservative, a sensitizer, an inorganic salt, and a surfactant, and the pH value of the pretreatment reagent is 7.2-7.6; the antibody latex reagent includes a buffer, latex particles combined with anti-human matrix metalloproteinase-3 antibodies, a preservative, and a protective agent, and the latex particles have a diameter of 100-400nm; the calibration reagent includes a quantitative matrix metalloproteinase-3 antigen. By combining the anti-human matrix metalloproteinase-3 antibody with the latex particles, the latex enhanced immunoturbidimetry method can be used to determine the content of MMP-3 in human serum, which has high accuracy, good repeatability, and is suitable for high-throughput detection. However, the patent does not focus on long-term stability.

[0005] Patent (CN115032402A) discloses a matrix metalloproteinase-3 latex detection kit, which is composed of the following raw materials: reaction solution 1, reaction solution 2 and matrix metalloproteinase-3 calibrator; reaction solution 1 includes the following components: 95-100g buffer, 0.9-1.8g electrolyte, 0.5-1g sensitizer, 0.1-0.5g stabilizer, 0.2-0.8g anti-interference agent and 0.2-0.4g antibacterial agent; reaction solution 2 is 10-20mL storage solution containing latex particles with surface-coupled matrix metalloproteinase-3 monoclonal antibody, and the matrix metalloproteinase-3 monoclonal antibody is sheep monoclonal antibody and / or rabbit monoclonal antibody; the mass fraction of latex particles in the storage solution is 0.1%-2%. The prepared kit has strong specificity, wide linear range, high sensitivity, high RF anti-interference performance and long-term stability. However, the opening stability and long-term stability of the patent still need to be improved.

[0006] Based on this, it is necessary to provide a matrix metalloproteinase-3 detection kit based on latex immunoturbidimetry that has both high sensitivity and high stability. Summary of the invention

[0007] The present invention aims at the problems existing in the prior art and provides a matrix metalloproteinase-3 detection kit based on latex immunoturbidimetry. The present invention adds a composite sensitizer consisting of dextran, polyethylene glycol 4000 and tributyl methyl ammonium chloride to reagent R1, and adds a composite stabilizer consisting of lithium chloride, hydroxylamine hydrochloride and a protective agent to reagent R2, and improves the detection sensitivity and the dispersion stability of latex particles through the different action mechanisms of each component, thereby providing a matrix metalloproteinase-3 detection kit with high sensitivity and high stability.

[0008] To achieve the above-mentioned purpose, in the first aspect, the present invention provides a matrix metalloproteinase-3 detection kit based on latex immunoturbidimetry, comprising: reagent R1 and reagent R2; the reagent R1 comprises: 1-5g / L sensitizer, 1-2g / L preservative, 0.1-1.0g / L chelating agent and 20-100mmol / L buffer 1; the reagent R2 is obtained by mixing latex particles coated with sheep anti-human matrix metalloproteinase-3 polyclonal antibodies and storage solution in a volume ratio of 1:3-15.

[0009] In a preferred embodiment, in the reagent R2, the content of latex particles coated with goat anti-human matrix metalloproteinase-3 polyclonal antibodies is 2-5 g / L and the particle size is 100-300 nm.

[0010] In a preferred embodiment, in reagent R2, the storage solution comprises, by weight percentage: 2-8% stabilizer, 0.5-2% inorganic salt, 0.1-0.5% surfactant and 20-100 mmol / L buffer 2 as the balance.

[0011] In a preferred embodiment, the sensitizer is a mixture of dextran, polyethylene glycol 4000 and tributylmethylammonium chloride in a mass ratio of 0.5-2:1:0.2-0.5.

[0012] The combined use of dextran, polyethylene glycol 4000 and tributyl methyl ammonium chloride can change the conformation of MMP-3 protein and keep it stable, thereby improving the detection sensitivity.

[0013] In a preferred embodiment, the preservative comprises at least one of sodium azide, ProClin 300 and methylisothiazolinone.

[0014] In a preferred embodiment, the complexing agent includes at least one of disodium ethylenediaminetetraacetate, dimercaptopropanol, sodium dimercaptopropane sulfonate, mercaptoethylamine, thioglycolic acid, and 8-hydroxyquinoline.

[0015] In a preferred embodiment, the buffer 1 is a phosphate buffer, preferably a buffer of sodium dihydrogen phosphate dihydrate-disodium hydrogen phosphate dodecahydrate.

[0016] In a preferred embodiment, the pH of buffer solution 1 and buffer solution 2 are each independently selected from 6.7-7.5.

[0017] In a preferred embodiment, the stabilizer is lithium chloride, hydroxylamine hydrochloride and a protective agent in a mass ratio of 0.5-2:1:1-3; the protective agent includes at least one of bovine serum albumin, sucrose, trehalose, glucose, mannitol, ethylene glycol and glycerol; more preferably, the protective agent is bovine serum albumin.

[0018] LiCl can provide Li + The dispersion stability of latex particles is improved through adsorption, solvation, etc., while hydroxylamine hydrochloride has an antioxidant effect. Combined with protective agents such as sugar, alcohol or bovine serum albumin, it can effectively improve the dispersibility and stability of latex particles.

[0019] In a preferred embodiment, the inorganic salt comprises sodium chloride and / or potassium chloride.

[0020] In a preferred embodiment, the surfactant comprises Tween-20 and / or Triton X-100.

[0021] In a preferred embodiment, the buffer 2 comprises one of Good's buffer, Tris buffer and phosphate buffer.

[0022] In a preferred embodiment, the matrix metalloproteinase-3 detection kit based on latex immunoturbidimetry also includes a calibrator and / or a quality control substance.

[0023] In a preferred embodiment, the calibrator comprises: recombinant human matrix metalloproteinase-3, bovine albumin and phosphate buffer solution; wherein the content of recombinant human matrix metalloproteinase-3 is 20-1600 ng / mL, the content of bovine albumin is 20-100 g / L, and the concentration of phosphate buffer solution is 20-100 mmol / L.

[0024] In a preferred embodiment, the quality control product includes quality control product 1 and quality control product 2.

[0025] In a preferred embodiment, the quality control product 1 includes: recombinant human matrix metalloproteinase-3, bovine albumin and phosphate buffer solution; wherein the content of recombinant human matrix metalloproteinase-3 is 50-450 ng / mL, the content of bovine albumin is 20-100 g / L, and the concentration of phosphate buffer solution is 20-100 mmol / L.

[0026] In a preferred embodiment, the quality control product 2 includes: recombinant human matrix metalloproteinase-3, bovine albumin and phosphate buffer solution; wherein the content of recombinant human matrix metalloproteinase-3 is 450-1600 ng / mL, the content of bovine albumin is 20-100 g / L, and the concentration of phosphate buffer solution is 20-100 mmol / L.

[0027] In a second aspect, the present invention provides the use of the aforementioned matrix metalloproteinase-3 detection kit based on latex immunoturbidimetry in the detection of matrix metalloproteinase-3.

[0028] The beneficial effects of the present invention are:

[0029] The present invention adds a composite sensitizer consisting of dextran, polyethylene glycol 4000 and tributyl methyl ammonium chloride to reagent R1, and adds a composite stabilizer consisting of lithium chloride, hydroxylamine hydrochloride and a protective agent to reagent R2, and improves the detection sensitivity and the dispersion stability of latex particles through the different action mechanisms of each component, thereby providing a matrix metalloproteinase-3 detection kit with both high sensitivity and high stability. DETAILED DESCRIPTION

[0030] The present invention will be further described below in conjunction with specific embodiments, and the advantages and features of the present invention will become clearer as the description proceeds. However, these embodiments are exemplary only and do not constitute any limitation to the scope of the present invention. It should be understood by those skilled in the art that the details and forms of the technical solution of the present invention may be modified or replaced without departing from the spirit and scope of the present invention, but these modifications and replacements all fall within the scope of protection of the present invention.

[0031] It is worth noting that the raw materials used in the present invention are all common commercially available products, and their sources are not specifically limited.

[0032] Among them, latex particles coated with sheep anti-human matrix metalloproteinase-3 polyclonal antibody were purchased from Shaoxing Chuangye Biotechnology Co., Ltd. with a specification of ≥20 g / L;

[0033] Dextran: T-40, purchased from Shanghai Zeye Biotechnology Co., Ltd.;

[0034] The remaining raw materials are commercially available.

[0035] Example 1

[0036] The components of reagent R1 are: sensitizer (a mixture of dextran, polyethylene glycol 4000 and tributyl methyl ammonium chloride in a mass ratio of 1:1:0.3) 3 g / L, preservative (sodium azide) 1.5 g / L, complexing agent (disodium ethylenediaminetetraacetic acid) 0.5 g / L and buffer 1 (100 mmol / L sodium dihydrogen phosphate dihydrate-disodium hydrogen phosphate dodecahydrate buffer, pH 7.4);

[0037] Reagent R2:

[0038] The storage solution is composed of the following by weight percentage: 5% of stabilizer (lithium chloride, hydroxylamine hydrochloride and bovine serum albumin in a mass ratio of 1:1:2), 1.2% of inorganic salt (potassium chloride), 0.3% of surfactant (Tween-20) and the balance of buffer 2 (50 mmol / L sodium dihydrogen phosphate dihydrate-sodium hydrogen phosphate dodecahydrate buffer, pH 7.4);

[0039] The preparation process of reagent R2 is:

[0040] The latex particles coated with goat anti-human matrix metalloproteinase-3 polyclonal antibody were mixed with the storage solution at a volume ratio of 1:5 to obtain reagent R2.

[0041] Example 2

[0042] The components of reagent R1 are: sensitizer (a mixture of dextran, polyethylene glycol 4000 and tributyl methyl ammonium chloride in a mass ratio of 0.5:1:0.2) 5 g / L, preservative (sodium azide) 1 g / L, complexing agent (disodium ethylenediaminetetraacetic acid) 0.2 g / L and buffer 1 (20 mmol / L sodium dihydrogen phosphate dihydrate-disodium hydrogen phosphate dodecahydrate buffer, pH 7.4);

[0043] Reagent R2:

[0044] The storage solution is composed of the following by weight percentage: 8% of stabilizer (lithium chloride, hydroxylamine hydrochloride and glycerol in a mass ratio of 0.5:1:3), 0.5% of inorganic salt (potassium chloride), 0.5% of surfactant (Triton X-100) and the balance of buffer 2 (20 mmol / L sodium dihydrogen phosphate dihydrate-sodium hydrogen phosphate dodecahydrate buffer, pH 7.4);

[0045] The preparation process of reagent R2 is the same as that of Example 1.

[0046] Example 3

[0047] The components of reagent R1 are: sensitizer (a mixture of dextran, polyethylene glycol 4000 and tributyl methyl ammonium chloride in a mass ratio of 2:1:0.5) 2 g / L, preservative (sodium azide) 2 g / L, complexing agent (disodium ethylenediaminetetraacetic acid) 1.0 g / L and buffer 1 (50 mmol / L sodium dihydrogen phosphate dihydrate-disodium hydrogen phosphate dodecahydrate buffer, pH 7.4);

[0048] Reagent R2:

[0049] The storage solution is composed of the following by weight percentage: 3% of a stabilizer (lithium chloride, hydroxylamine hydrochloride and glucose in a mass ratio of 2:1:1), 2% of an inorganic salt (sodium chloride), 0.1% of a surfactant (Tween-20) and the remainder of buffer 2 (100 mmol / L of a buffer of sodium dihydrogen phosphate dihydrate-sodium hydrogen phosphate dodecahydrate, pH 7.4);

[0050] The preparation process of reagent R2 is the same as that of Example 1.

[0051] Comparative Example 1

[0052] Except that hydroxylamine hydrochloride is used in place of lithium chloride in the stabilizer, the rest is the same as in Example 1.

[0053] Comparative Example 2

[0054] Except that lithium chloride is used in place of hydroxylamine hydrochloride in the stabilizer, the rest is the same as in Example 1.

[0055] Comparative Example 3

[0056] Except that bovine serum albumin is used in place of hydroxylamine hydrochloride in the stabilizer, the rest is the same as in Example 1.

[0057] Comparative Example 4

[0058] Except that polyethylene glycol 6000 is used instead of polyethylene glycol 4000 in the sensitizer, the rest is the same as Example 1.

[0059] Comparative Example 5

[0060] The same procedures as in Example 1 were followed except that the sensitizer was a mixture of dextran, polyethylene glycol 4000 and tributylmethylammonium chloride in a mass ratio of 0.3:1:1.

[0061] Comparative Example 6

[0062] The same procedures as in Example 1 were followed except that the sensitizer was a mixture of dextran, polyethylene glycol 4000 and tributylmethylammonium chloride in a mass ratio of 1:0.3:1.

[0063] Application Examples

[0064] The reagents of Examples 1-3 and Comparative Examples 1-6 were used in a matrix metalloproteinase-3 detection kit based on latex immunoturbidimetry, and performance tests and evaluations were performed. The test instrument was a Hitachi biochemical analyzer (7180). The test conditions were shown in Table 1, and the determination steps were shown in Table 2:

[0065] Table 1 Test conditions

[0066] temperature 37℃ Reaction direction Rising (+) Dominant wavelength 700nm Incubation time / reaction time 3-5min / 5min Calibration Method Multi-point calibration S / R1 / R2 3 / 150 / 50μL optical path 1.0cm Calibration Type Spline

[0067] Table 2 Determination steps

[0068]

[0069] Among them, standards of different concentrations were prepared using recombinant human matrix metalloproteinase-3 (contents of 100.05 ng / mL, 200.10 ng / mL, 400.20 ng / mL, 800.40 ng / mL, and 1600.80 ng / mL, respectively), bovine albumin (50 g / L), and phosphate buffer solution (pH 7.4, 0.05 mol / L), and quality control products 1 and 2 were prepared using recombinant human matrix metalloproteinase-3 (contents of 134.90 ng / mL and 589.10 ng / mL, respectively), bovine albumin (50 g / L), and phosphate buffer solution (pH 7.4, 0.05 mol / L), respectively.

[0070] (1) Reagent R2 should appear as a milky white solution without any precipitation.

[0071] (2) Reagent blank absorbance: At the corresponding dominant wavelength (700 nm), the reagent blank absorbance (A) is ≤1.5.

[0072] (3) Analytical sensitivity: When the test sample concentration is 800 ng / mL, the absorbance difference ΔA ≥ 0.0100.

[0073] (4) Linear range: The reagent is within the specified linear interval (20-1600 ng / mL): Calculate the linear correlation coefficient (r), generally r ≥ 0.990.

[0074] (5) Batch variation: When reagents from different batches are used to test the same sample, the relative range (R) between batches is calculated. Generally, R is ≤ 10.0%. The smaller the R, the smaller the batch variation.

[0075] (6) Precision: Repeat the test on a sample 10 times and calculate the coefficient of variation (CV). Generally, CV is ≤ 8%. The smaller the CV, the better the precision.

[0076] (7) Open bottle stability: After opening the test kit, place it in the reagent compartment of a biochemical analyzer (2-8°C) for 49 days. Test the quality control product every 7 days, 3 times each time, and calculate the relative deviation between the average value and the value on day 0, i.e., relative deviation = (average value on day n - average value on day 0) / average value on day 0 × 100%, which should generally be ≤10.0.

[0077] (8) Thermally accelerated stability: The reagents were placed at 37°C for 8 days. The reagents were then taken out and the appearance of reagent R2, reagent blank absorbance, analytical sensitivity, linear range, batch difference and precision were evaluated.

[0078] (9) Long-term stability: The reagent was placed at 2-8°C for 28 months. The properties of reagent R2, reagent blank absorbance, analytical sensitivity, linear correlation coefficient, batch difference and precision were evaluated every 7 months.

[0079] 1. The reagent performance of the embodiment and the comparison group at month 0 (initial state) is shown in Table 3:

[0080] Table 3 Performance data of the embodiments and comparative examples at month 0

[0081]

[0082] It can be seen from Table 3 that in the initial state, the shape, blank absorbance, sensitivity, linear correlation coefficient, batch difference and precision of the reagent R2 of Examples 1-3 are all good and can meet the requirements. Comparative Examples 1-6 are inferior to Examples 1-3 in different indicators, especially the analytical sensitivity of Comparative Examples 5-6 does not meet the requirements.

[0083] 2. The results of stability after opening the bottle are shown in Table 4:

[0084] Table 4 Opening stability of the embodiments and comparative examples (quality control product 1)

[0085]

[0086]

[0087] As can be seen from Table 4, the reagents of Examples 1-3 were opened on the machine for 49 days, and the maximum deviation of the measured values ​​compared with the 0th day was 2.03%, and the opening of the bottle was stable. For the quality control product 2 with a higher concentration, the change trend of the measured values ​​after opening the bottle of Examples 1-3 and Comparative Examples 1-6 was similar to Table 4, and the overall degree of change was slightly lower than that of the quality control product 1. It can be seen that the reagents of Examples 1-3 have good opening stability, and the defined stabilizers and sensitizers can improve the opening stability.

[0088] 3. The results of thermal acceleration stability are shown in Table 5:

[0089] Table 5 Summary of performance of embodiments and comparative examples before and after thermal acceleration test

[0090]

[0091]

[0092] As can be seen from Table 5, Examples 1-3 have excellent thermal stability. After the thermal acceleration test, the properties of R2, reagent blank absorbance, analytical sensitivity, linear correlation coefficient, batch difference and precision did not change much, while Comparative Examples 1-6 had obvious changes.

[0093] 4. Long-term stability results are shown in Table 6-11:

[0094] Table 6 Properties of R2 reagents in Examples and Comparative Examples

[0095] Group Month 0 Month 7 Month 14 Month 21 Month 28 Example 1 No precipitation No precipitation No precipitation No precipitation No precipitation Example 2 No precipitation No precipitation No precipitation No precipitation No precipitation Example 3 No precipitation No precipitation No precipitation No precipitation No precipitation Comparative Example 1 No precipitation No precipitation There is precipitation There is precipitation There is precipitation Comparative Example 2 No precipitation No precipitation There is precipitation There is precipitation There is precipitation Comparative Example 3 No precipitation There is precipitation There is precipitation There is precipitation There is precipitation Comparative Example 4 No precipitation No precipitation No precipitation No precipitation No precipitation Comparative Example 5 No precipitation No precipitation No precipitation No precipitation No precipitation Comparative Example 6 No precipitation No precipitation No precipitation No precipitation No precipitation

[0096] It can be seen from Table 6 that the R2 reagents of Examples 1-3 are very stable, while changing the stability composition of the R2 reagents in Comparative Examples 1-3 will greatly affect the long-term stability of the R2 reagents, and precipitation will occur after being placed for 7-14 months.

[0097] Table 7 Long-term stability of blank absorbance of examples and comparative examples

[0098]

[0099]

[0100] It can be seen from Table 7 that after long-term storage, the reagent blank absorbance of Examples 1-3 is basically stable, while changing the composition of the stability or sensitizer in Comparative Examples 1-6 will cause the reagent blank absorbance to change significantly after long-term storage.

[0101] Table 8 Long-term stability of analytical sensitivity of embodiments and comparative examples

[0102] Group Month 0 Month 7 Month 14 Month 21 Month 28 Example 1 0.2562 0.2525 0.2518 0.2509 0.2497 Example 2 0.2535 0.2511 0.2503 0.2493 0.2488 Example 3 0.2546 0.2518 0.2509 0.2496 0.2490 Comparative Example 1 0.2438 0.1756 0.1241 0.0756 0.0291 Comparative Example 2 0.2410 0.1718 0.1234 0.0745 0.0289 Comparative Example 3 0.2465 0.1803 0.1382 0.0839 0.0315 Comparative Example 4 0.1568 0.1231 0.0913 0.0431 0.0095 Comparative Example 5 0.0804 0.0545 0.0198 0.0065 0.0032 Comparative Example 6 0.0568 0.0379 0.0133 0.0048 0.0025

[0103] It can be seen from Table 8 that the analytical sensitivity of Examples 1-3 does not change much after long-term storage, while in Comparative Examples 1-6, the analytical sensitivity obviously deteriorates with the storage time when the stability or sensitizer composition is changed.

[0104] Table 9 Long-term stability of linear correlation coefficients of embodiments and comparative examples

[0105] Group Month 0 Month 7 Month 14 Month 21 Month 28 Example 1 0.9993 0.9991 0.9992 0.9990 0.9994 Example 2 0.9987 0.9985 0.9983 0.9980 0.9982 Example 3 0.9995 0.9994 0.9992 0.9992 0.9986 Comparative Example 1 0.9957 0.9932 0.9941 0.9925 0.9923 Comparative Example 2 0.9962 0.9945 0.0023 0.9937 0.9914 Comparative Example 3 0.9938 0.9931 0.9926 0.9915 0.9903 Comparative Example 4 0.9948 0.9940 0.9936 0.9924 0.9911 Comparative Example 5 0.9954 0.9946 0.9951 0.9938 0.9930 Comparative Example 6 0.9961 0.9952 0.9940 0.9928 0.9931

[0106] Table 10 Batch Difference (%) of Examples and Comparative Examples Long-term Stability

[0107]

[0108]

[0109] Table 11 Precision (%) Long-term stability of examples and comparative examples

[0110] Group Month 0 Month 7 Month 14 Month 21 Month 28 Example 1 2.77 3.48 3.86 2.73 3.09 Example 2 2.56 2.85 3.64 3.78 3.25 Example 3 2.96 3.58 4.04 3.10 3.63 Comparative Example 1 5.93 8.76 7.54 10.12 12.34 Comparative Example 2 6.31 7.99 9.02 11.89 13.45 Comparative Example 3 5.98 8.35 10.78 9.32 12.63 Comparative Example 4 3.44 5.77 7.98 9.81 10.33 Comparative Example 5 4.23 6.32 6.89 9.03 11.45 Comparative Example 6 4.10 8.21 7.47 10.23 12.67

[0111] It can be seen from Tables 9-11 that after long-term storage, the linear correlation coefficient, batch difference and precision of Examples 1-3 did not change much, while in Comparative Examples 1-6, when the composition of the stability or sensitizer was changed, the batch difference and precision both decreased with the storage time, which did not meet the requirements.

[0112] In summary, by using the defined stabilizers and sensitizers, the reagents of Examples 1-3 can be used to prepare a matrix metalloproteinase-3 detection kit based on latex immunoturbidimetry with high sensitivity and high stability.

[0113] The solution of the present invention is not limited to the technical means disclosed by the above technical means, but also includes a technical solution composed of any combination of the above technical features. The above is a specific implementation of the present invention. It should be pointed out that for ordinary technicians in this field, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications are also considered as the protection scope of the present invention.

Claims

1. A matrix metalloproteinase-3 detection kit based on latex immunoturbidimetry, characterized in that: include: Reagent R1 and reagent R2; The reagent R1 comprises: 1-5 g / L of sensitizer, 1-2 g / L of preservative, 0.1-1.0 g / L of complexing agent and 20-100 mmol / L of buffer 1; the reagent R2 is obtained by mixing latex particles coated with sheep anti-human matrix metalloproteinase-3 polyclonal antibody and storage solution in a volume ratio of 1:3-15; the sensitizer is dextran, polyethylene glycol 4000 and tributyl ether in a mass ratio of 0.5-2:1:0.2-0.5 The storage solution comprises, by weight percentage, 2-8% of a stabilizer, 0.5-2% of an inorganic salt, 0.1-0.5% of a surfactant and a balance of 20-100 mmol / L buffer 2; the stabilizer is lithium chloride, hydroxylamine hydrochloride and a protective agent in a mass ratio of 0.5-2:1:1-3; the protective agent comprises at least one of bovine serum albumin, sucrose, trehalose, glucose, mannitol, ethylene glycol and glycerol.

2. The matrix metalloproteinase-3 detection kit according to claim 1, characterized in that: In reagent R2, the content of latex particles coated with sheep anti-human matrix metalloproteinase-3 polyclonal antibody is 2-5 g / L and the particle size is 100-300 nm.

3. The matrix metalloproteinase-3 detection kit according to claim 1, characterized in that: The preservative includes at least one of sodium azide, ProClin 300 and methylisothiazolinone; the chelating agent includes at least one of disodium ethylenediaminetetraacetate, dimercaptopropanol, sodium dimercaptopropane sulfonate, mercaptoethylamine, thioglycolic acid and 8-hydroxyquinoline; and the buffer 1 is a phosphate buffer.

4. The matrix metalloproteinase-3 detection kit according to claim 3, characterized in that: The buffer solution 1 is a buffer solution of sodium dihydrogen phosphate dihydrate-sodium hydrogen phosphate dodecahydrate.

5. The matrix metalloproteinase-3 detection kit according to claim 1, characterized in that: The inorganic salt includes sodium chloride and / or potassium chloride; the surfactant includes Tween-20 and / or Triton X-100.

6. The matrix metalloproteinase-3 detection kit according to claim 1, characterized in that: The buffer 2 includes one of Good's buffer, Tris buffer and phosphate buffer.

7. The matrix metalloproteinase-3 detection kit according to claim 1, characterized in that: The matrix metalloproteinase-3 detection kit also includes calibrators and / or quality control products.

8. Use of the matrix metalloproteinase-3 detection kit based on latex immunoturbidimetry according to any one of claims 1 to 7 in the detection of matrix metalloproteinase-3.

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