IgG type rheumatoid factor enzyme immunity detection method

Abstract

The invention discloses an IgG type rheumatoid factor enzyme immunity detection method. The method comprises the following steps of: (1) preparing reagents and specimens; (2) adding IgG-RF standard products, specimens and quality control products into coated plate micropores; (3) under the action of a certain temperature, pouring off liquid in the micropores, washing and beating to dry; (4) adding enzyme markers into all the micropores; (5) repeating the step (3); (6) adding substrates into all the micropores; (7) adding stopping solutions into all the micropores and uniformly shaking; (8) reading a light-sucking value A by using a microplate reader; and (9) calculating, wherein the coating plate micropores in the step (2) are IgG-Fc segment coated plate micropores, and the enzyme markers in the step (4) are anti-IgG-F(ab1)2 segment antibodies. The IgG type rheumatoid factor enzyme immunity detection method eliminates the influence of a cross reaction, greatly improves the result accuracy, and obtains a good result proved by clinical applications.

Description

technical field [0001] The invention relates to an enzyme immunoassay method for rheumatoid factor typing, in particular to an IgG type rheumatoid factor enzyme immunoassay method. Background technique [0002] Rheumatoid factor (rheumatoid factor, referred to as RF) is an anti-IgG antibody, which can react with certain regions on the antigenic determinant, that is, IgG-Fc fragment, so it is also called anti-IgG-Fc fragment antibody. [0003] There are five types of rheumatoid factor according to Ig type, namely IgM-RF, IgG-RF, IgA-RF, IgD-RF, IgE-RF. As an autoantibody, rheumatoid factor, especially IgM-RF and IgG-RF, can specifically combine with antigen IgG in vivo to form an antigen-antibody complex---immune complex (IC for short), if it is not cleared by the body in time, That is, the circulating immune complex (CIC) is formed in the blood circulation in the body, and under certain conditions, it is easy to deposit on the synovial membrane, superficial surface and bloo...

AI Technical Summary

Problems solved by technology

However, IgG-RF cannot be detected by agglutination test because IgG is a monomeric molecule. At the same time, because both the antigen and the tested IgG-RF are IgG, general detection techniques cannot distinguish antigen and IgG-RF. Therefore, IgG-RF is usually detected by enzyme Immunization technology (ELISA for short), uses rabbit IgG that has germline differences from normal human IgG or human denatured human IgG that denatures human IgG by heating to cause heterogeneity as the coating antigen, and then uses enzyme-labeled anti-human IgG as the second Secondary antibody to detect IgG-RF

This method aims to eliminate the interference of IgG as an antigen on the IgG-RF to be detected, but there are still cross-reaction effects and the accuracy of the results

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