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Complement fixation anti-sperm antibody enzyme-linked immunosorbent assay method

An enzyme-linked immunosorbent assay and anti-sperm antibody technology, applied in the field of enzyme immunoassay

Inactive Publication Date: 2016-11-09
TIANJIN BAODI HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the detection of IgG, IgM, IgA, IgD, and IgE total antisperm antibodies by complement-mediated enzyme-linked immunoassay at home and abroad.

Method used

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  • Complement fixation anti-sperm antibody enzyme-linked immunosorbent assay method
  • Complement fixation anti-sperm antibody enzyme-linked immunosorbent assay method
  • Complement fixation anti-sperm antibody enzyme-linked immunosorbent assay method

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Experimental program
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Embodiment 1

[0011] 1. Preparation of anti-sperm antibody ELISA plate

[0012] Dilute the sperm antigen with the coating solution to 1:200 times, add 100 μl to the reaction wells of the coated microwell reaction plate, add 350 μl / each well of phosphate buffer containing 1% bovine serum albumin, block at room temperature for 2 hours, 4 ℃ overnight. After shaking off the liquid in the hole, wash it 3 times with washing solution for 3 minutes each time (commercially available).

[0013] 2. Preparation of enzyme-labeled antibody

[0014] (1) Dissolve 5 mg of horseradish peroxidase in 0.5 ml of 0.2 mol / L pH5.6 HAC-NaAc buffer solution, and drop in 0.1 mol / L NaIO 4 0.25ml, 30 minutes at 4°C.

[0015] (2) Add 0.5ml of 2.5% ethylene glycol, 30min at room temperature.

[0016] (3) Add anti-complement C 3C Antibody 5 ~ 10mg, adjust the pH of the reaction solution to about 9.0 with 1.0mmol / L pH9.5 carbonate buffer solution, overnight at 4°C (commercially available, complement C 3C commercially ...

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Abstract

The invention discloses a complement fixation anti-sperm antibody enzyme-linked immunosorbent assay method and belongs to the technical field of enzyme immunoassay. The method comprises the following steps: combining IgG, IgA, IgM, IgD and IgE type anti-sperm antibodies in serum with a reaction hole coated sperm antigen, then combining with a complement C3C to form antigen-antibody-complement C3C immune complex, adding anticomplement C3C labelling enzyme, reacting, then adding an enzyme substrate solution, and developing. The method disclosed by the invention can detect the IgG, IgA, IgM, IgD and IgE type anti-sperm antibodies in the serum at the same time, is easy to operate, rapid, sensitive, accurate and cheap, is applicable to total anti-sperm antibody detection and can be taken as an important examination index of infertility.

Description

technical field [0001] The invention relates to an enzyme-linked immunosorbent detection method for complement fixation in serum, which belongs to the technical field of enzyme immunoassay. Background technique [0002] Anti-sperm antibody (AsAb) is a complex pathological product that can affect both men and women. The exact cause is not yet fully understood. Men’s sperm and seminal plasma are all specific antigens for women. After contact with blood, both men and women can Cause immune reaction, produce corresponding antibody, prevent sperm from combining with egg, and cause infertility, in the blood and seminal plasma of 5-10% infertile men, sperm agglutinating antibody and sperm braking antibody do exist. When the serum and seminal plasma with anti-sperm antibodies are mixed with normal sperm, the sperm will undergo head-to-head, tail-to-tail or mixed agglutination, or in the presence of complement, this serum and seminal plasma can make swimming Sperm stops moving or sh...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
CPCG01N33/543
Inventor 李立和丁弘张超
Owner TIANJIN BAODI HOSPITAL
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