ELISA (enzyme-linked immunosorbent assay) kit for EV (enterovirus) 71 inactivated vaccine antigen

Abstract

The invention relates to an ELISA (enzyme-linked immunosorbent assay) kit for an EV (enterovirus) 71 inactivated vaccine antigen. The detection kit comprises an EV71 pre-coated polyclonal antibody ELISA plate, a sample diluent, a second antibody, an enzyme-labeled antibody, a concentrated cleaning solution, an enzyme substrate solution and a stop buffer, wherein the ELISA plate is pre-coated with a polyclonal antibody prepared by taking recombinant EV71 structural protein 1 as an immune source; the second antibody adopts a monoclonal antibody prepared by taking keyhole limpet hemocyanin coupled polypeptide sequence FGEHKQEKDL as the immune source; the enzyme-labeled antibody adopts a horse radish peroxidase labelled goat anti-mouse immunoglobulin antibody; and an antibody standard is placed in the kit. The ELISA kit has better sensitivity when measuring the titer of the EV71 inactivated vaccine antigen, and has better linearity in the range of 5.9-750 ng / ml, and the linearly dependent coefficient r2 is larger than 0.99. The kit for measuring the titer of the EV71 inactivated vaccine antigen simply and conveniently has good specificity, accurate quantification, high sensitivity and good repeatability.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to an ELISA detection kit for enterovirus 71 (EV71) inactivated vaccine antigen. Background technique [0002] In recent years, hand, foot and mouth disease has ravaged most parts of our country and brought great harm to people. A large number of epidemiological studies have shown that enterovirus 71 is one of the main pathogens causing HFMD. EV71 belongs to the Picornaviridae family, a member of the Enterovirus genus. The infection of this virus mostly occurs in infants under 5 years old, and can cause herpes on the hands, feet, mouth and other parts. Individual patients can cause complications such as myocarditis, pulmonary edema, and aseptic meningoencephalitis. morbidity and mortality rates. Therefore, it is of great significance to develop a safe and effective EV71 vaccine to prevent and control the spread of HFMD. At present, there is no related vaccine in the market, and the ...

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Problems solved by technology

However, this method has a long cycle and often takes several days to get the results, and because CPE can only detect infectious viruses, it cannot correctly detect the antigen content in inactivated vaccines, such as vacuolar viruses, inactivated viruses etc., so it is impossible to carry out effective, real-time and correct monitoring during the production process of the vaccine

However, the detection of EV71 antigen by enzyme-linked immunosorbent assay often has a certain impact on the detection results due to the source of the antibody, especially the non-specific adsorption of impurities in the vaccine greatly interferes with the detection results.

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