A kind of swine fever virus blocking ELISA antibody detection kit and its application

Abstract

The invention discloses a hog cholera virus inhibition ELISA antibody detection kit and application thereof. The kit comprises hog cholera virus E2 protein which is expressed by virtue of a baculovirus expression system and a cell suspension culture process; the hog cholera virus E2 protein is purified to serve as a coated plate antigen and an immunogen to prepare a monoclonal antibody of the hog cholera virus E2 protein, and horse radish peroxidase (a hybridoma cell strain 4A7 CCTCC NO.C2014229) is marked. The kit further comprises an antigen coated plate, a positive control, a negative control, sample diluent, a scrubbing solution, a developing solution A, a developing solution B and a stop solution. According to the application of the hog cholera virus inhibition ELISA antibody detection kit in the detection of the hog cholera virus antibody, the kit is used for detecting other positive serums except hog cholera viruses, the cross reaction is avoided, the specificity is strong, the sensitivity is high, the detection time is short, and the repeatability is good; compared with an import kit, the detection coincidence rate reaches above 95%.

Description

technical field [0001] The present invention relates to the detection of animal epidemics, in particular to an enzyme-linked immunoimmunoassay kit capable of large-scale production for detecting antibodies against swine fever virus, and at the same time relates to a detection kit for detecting antibodies against swine fever virus by ELISA in the application. It is suitable for detecting the antibody level of swine fever virus in pig serum. Background technique [0002] Classical Swine Fever (CSF) is one of the infectious diseases that seriously endanger the pig industry. It is highly contagious and highly pathogenic, and has caused huge economic losses to the pig industry worldwide. It has been recognized by the World Organization for Animal Health (OIE ) is listed as one of the 16 infectious diseases in category A, and it is an animal infectious disease that must be reported. my country also lists it as a first-class infectious disease. In recent years, the symptoms of swi...

AI Technical Summary

Problems solved by technology

At present, most of the ELISA methods for detecting swine fever antibodies established in China are indirect ELISA methods. Although the indirect ELISA method for detecting swine fever antibodies appeared earlier and has good sensitivity, it is often inevitable that there will be high non-specificity.

Secondly, most of the coating antigens used in the established ELISA method for detecting classical swine fever antibodies are expressed in prokaryotic systems, but the structural protein E2 of classical swine fever virus is a glycosylated protein, and the prokaryotic expression products cannot be glycosylated and other proteins. Modification, it is quite different from the natural E2 protein in terms of conformation and reactogenicity

[0004] The blocking ELISA kit for the detection of classical swine fever virus antibody that the inventor has previously published (published in the Proceedings of the Eighth National Member Congress and the Fifteenth Academic Symposium of the Animal Infectious Diseases Branch of the Chinese Association of Animal Husbandry and Veterinary Medicine, 2013.10) Compared with imported kits of the same kind, there are still shortcomings such as low negative coincidence rate, poor repeatability, and long detection time, so it is necessary to improve the antigen preparation process, monoclonal antibody preparation, and kit reaction conditions. Work

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