Preparation method of recombinant protein IE1-coated ELISA (Enzyme linked immune sorbent assay) reaction plate and assay kit for quantitatively detecting HCMV (human cytomegalovirus) neutralized antibody in human plasma

Abstract

The invention discloses a preparation method of a recombinant protein IE1-coated ELISA (Enzyme linked immune sorbent assay) reaction plate and an assay kit for quantitatively detecting HCMV (human cytomegalovirus) neutralized antibody in human plasma and relates to a preparation method of a novel recombinant antigen IE1-coated ELISA reaction plate and a prepared ELISA assay kit for quantitatively detecting HCMV neutralized antibody in human plasma based on the preparation method of the novel recombinant antigen IE1-coated ELISA reaction plate. The preparation method comprises the following steps: preparing specificity recombinant HCMVIE1 antigen, purifying protein, coating the ELISA reaction plate and assembling the ELISA assay kit for quantitatively detecting HCMV neutralized antibody in human plasma. Compared with the other commercial ELISA assay kit, the ELISA assay kit disclosed by the invention can highly specifically, sensitively and rapidly detect IgG antibody titer with neutralized HCMV specificity in plasma and the detection result is highly consistent with that of a micro neutralization experiment. The preparation method of the recombinant protein IE1-coated ELISA (Enzyme linked immune sorbent assay) reaction plate and the assay kit for quantitatively detecting HCMV (human cytomegalovirus) neutralized antibody in human plasma are mainly used for screening high-titer HCMV intravenous injected gamma globulin production raw materials for laboratory research, clinical screening and blood product enterprises screening.

Description

technical field [0001] The invention relates to the fields of genetic engineering technology, diagnostic reagents and research and development of blood products, in particular to a method for preparing a recombinant protein IE1-coated enzyme-labeled reaction plate and a kit for quantitatively detecting human plasma HCMV neutralizing antibodies. Background technique [0002] Human cytomegalovirus (HCMV) belongs to the subfamily of human herpesviruses, and its infection is very common in the population, with a worldwide distribution. Serological epidemiological surveys show that 40-100% of adults have HCMV antibodies. Current research shows that HCMV is one of the most common pathogens of human congenital infection, which has a huge impact on pregnant women and fetuses, often causing miscarriage, stillbirth, fetal malformation, growth retardation and delayed central nervous system defects and other diseases. [0003] Pregnant women during pregnancy (especially early pregnancy)...

AI Technical Summary

Problems solved by technology

Therefore, high-titer plasma screened with these kits cannot be used for the production of specific intravenous gamma globulin preparations

This is related to the coating antigen selected by the ELISA kit. HCMV-infected organisms are mainly induced by the pp150 protein to produce humoral immunity. Therefore, most commercially available ELISA kits use the pp150 protein as the coating antigen, but the antigen for the pp150 protein does not have Ability to neutralize viruses

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