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Matrix metalloproteinase-3(MMP3) detection kit

A detection kit and matrix metal technology, which are applied in the field of medical biological detection, can solve the problems of unfavorable clinical popularization, insufficient detection sensitivity, high cost and the like, and achieve the effects of low cost, high specificity and good stability.

Inactive Publication Date: 2016-09-28
河北艾驰生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, there are few kits for detecting matrix metalloproteinase-3, and the method also has major defects, such as insufficient sensitivity, cumbersome operation, high cost, etc., which are not conducive to clinical promotion and use.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: Preparation of anti-matrix metalloproteinase-3 antibody latex particles.

[0039] 1) Prepare 50mM MES buffer solution with a pH value of 6.0;

[0040] 2) Dilute the latex microspheres with the above-mentioned MES buffer solution, the concentration is 1% (v / v);

[0041] 3) Add EDAC to the latex microsphere suspension at a concentration of 40 mg / ml, and incubate at room temperature for 40 minutes;

[0042] 4) Wash the latex microspheres with the same volume of MES buffer, then centrifuge at 13000g / min for 30 minutes, wash the latex microspheres again with the same volume of MES buffer, then centrifuge at 13000g / min for 30 minutes, and then dissolve the latex microspheres in MES buffer;

[0043] 5) Dissolving the rabbit anti-human matrix metalloproteinase-3 antibody in the above-mentioned MES buffer at a concentration of 1 mg / mL;

[0044] 6) Mixing the antibody solution and the latex microsphere suspension, stirring and reacting at 37°C for 3 hours;

[0045]...

Embodiment 2

[0047] Embodiment 2: the preparation method of kit.

[0048] (1) Reagent 1:

[0049] Tris 100mM, sodium azide 5g / L, PEG-6000 4g / L, pH value 8.1;

[0050] (2) Reagent 2:

[0051]Tris 100mM, sodium azide 2g / L, methylparaben 1g / L, anti-human matrix metalloproteinase-3 antibody latex particles 1g / L, pH value 6.0;

[0052] (3) Calibrator:

[0053] Level 1 contains matrix metalloproteinase-3 antigen 37.5ng / ml, sodium azide 5g / L; level 2 contains matrix metalloproteinase-3 antigen 75ng / ml, sodium azide 5g / L; level 3 contains matrix metalloproteinase-3 antigen 150ng / ml, sodium azide 5g / L; level 4 containing matrix metalloproteinase-3 antigen 300ng / ml, sodium azide 5g / L;

[0054] (4) Quality control products:

[0055] Matrix metalloproteinase-3 antigen 50ng / ml, sodium azide 5g / L.

Embodiment 3

[0056] Embodiment 3: detection method.

[0057] The matrix metalloproteinase-3 assay kit described in this example uses an automatic biochemical analyzer with dual reagent functions, such as Hitachi 7180 automatic biochemical analyzer, etc., and uses the two-point endpoint method for determination. Put reagent 1 and reagent 2 on the reagent position of the instrument at a ratio of 3:1, and place purified water, calibrators and samples on the corresponding positions of the sample tray. The operation is as follows:

[0058] Table 1 Example 2 reagent detection method

[0059]

[0060]

[0061] Calculation: MMP3 content (ng / mL) = ΔA 测定 / ΔA 测定 ×C 标准

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Abstract

The invention relates to a matrix metalloproteinase-3(MMP3) detection kit and a detection method thereof, and belongs to the technical field of medical and biological detection. The kit comprises a first reagent, a second reagent, a calibrator and a quality control product, wherein the first reagent comprises Tris-HCl, sodium azide, and PEG-6000, and the second reagent comprises Tris-HCl, anti-human matrix metalloproteinase-3 antibody coated latex particles, sodium azide and methyl paraben. The calibrator comprises a matrix metalloproteinase-3 antigen and sodium azide, and the quality control product comprises a matrix metalloproteinase-3 antigen and sodium azide. The matrix metalloproteinase-3(MMP3) detection kit and the detection method thereof provided by the invention adopt a latex immune turbidimetric technology, has the advantages of high analytical sensitivity, accurate results, simple operation, suitable cost and the like, and is convenient for popularization.

Description

technical field [0001] The invention belongs to the technical field of medical biological detection, in particular to a matrix metalloproteinase-3 (MMP3) detection kit. Background technique [0002] Matrix metalloproteinases are a large family, so named because they require metal ions such as Ca2+ and Zn2+ as cofactors. Its family members have a similar structure and are generally composed of five structural domains with different functions: (1) the hydrophobic signal peptide sequence; (2) the propeptide region, whose main function is to maintain the stability of the zymogen. When this region is cleaved by exogenous enzymes, the MMPs zymogen is activated; (3) the catalytic active region has a zinc ion binding site, which is crucial for the enzyme's catalytic effect; (4) the proline-rich Hinge region; (5) carboxy-terminal region, which is related to the substrate specificity of the enzyme. Among them, the enzyme catalytic active region and the propeptide region are highly c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531
CPCG01N33/531
Inventor 张伟林黄龙梅王鑫
Owner 河北艾驰生物科技有限公司
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