An enzyme-linked immunosorbent assay kit for detecting the content of anti-Müllerian hormone and its detection method

Abstract

The invention provides a double-antibody sandwich enzyme-linked immunosorbent assay kit for detecting the content of anti-mullerian hormone and a detection method of the double-antibody sandwich enzyme-linked immunosorbent assay kit. The kit comprises an elisa plate coated with a capture antibody, a sample diluent, a standard substance, a biotin-labeled detection antibody, horseradish peroxidase-labeled avidin, a concentrated washing solution, a substrate solution and a stop solution, wherein the capture antibody is a mouse anti-human AMH antibody; the sample diluent is prepared from 1 * PBS,1% of BSA, 0.05% of Tween-20 and 0.1% of Proclin 300; the standard substance is a human AMH freeze-dried standard substance; the detection antibody is an anti-human AMH antibody; the concentrated washing liquid is 25 * PBST containing 25 * PBS, 1.25% of Tween 20 and 2.5% of Proclin 300, and the pH value of the concentrated washing liquid is 7.6; the substrate solution is a TMB chromogenic substrate solution; and the stop solution is 2M sulfuric acid. According to the kit and the detection method thereof, the manpower and equipment costs are reduced, the usage amount of raw materials and samples is reduced, and the cost is reduced, so that the use efficiency is improved.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to an ELISA kit for detecting the content of anti-Müllerian hormone and a detection method thereof. Background technique [0002] Anti-Müllerian hormone, also known as Muellerian-inhibiting factor (Anti-Muellerian hormone AMH), is a member of the transforming growth factor β (TGF-β) superfamily and is a of polypeptide hormones. The human AMH gene is located on the short arm of chromosome 19, contains 5 exons, and encodes a protein precursor of 560 amino acids. . AMH is a homodimeric glycoprotein composed of two monomers with a molecular weight of 70kDa linked by a disulfide bond. The activation of AMH requires hydrolysis to release the carboxy-terminal dimer, so as to have the biological activity of promoting the degeneration of Müllerian ducts And can inhibit the growth and differentiation of certain tumors. AMH and inhibins (Inhibins), activins (Activins), growt...

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Problems solved by technology

Chemiluminescence and electrochemiluminescence have high sensitivity but require expensive special equipment, which is not suitable for small batch detection or general laboratory use; colloidal gold method is simple to operate, short reaction time, easy to use but not suitable for accurate quantitative analysis; When the colloidal gold method is used in combination with the fluorescent immunochromatography method, there are more requirements for the test strips, and the processing process is troublesome; the latex-enhanced immunoturbidimetric method is simple to operate, quick in response and accurate in quantification, and can be used clinically, but its low The value repeatability is poor and a special biochemical analyzer is required, which is expensive

[0006] Enzyme-linked immunoassay uses 96-microwell plates as solid phase carriers to coat antibodies, and uses antigen-antibody immune reactions to achieve quantitative detection; it is widely used due to its advantages of good specificity, high sensitivity, accurate quantification, and low requirements for experimental equipment. , but it has the disadvantages of long reaction time, cumbersome steps, poor repeatability, etc.

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