Peste des petits ruminants virus H protein antibody iELISA detection method and application

Abstract

According to the invention, the B cell epitope of the screened peste des petits ruminants virus (PPRV) H protein is used as a basis, four antigen epitope peptides with better reactogenicity are selected and are connected in series to be synthesized into a coating antigen; an iELISA method for the PPRV H protein antibody is established, the optimal coating amount of the multi-epitope antigen peptide is 1.5*10<-6> mu g / hole, the negative and positive critical dilution concentration of to-be-detected serum is 1: 100, and the optimal dilution concentration of the enzyme-labeled secondary antibodyis 1: 80000; the coating solution is a carbonate buffer solution, the serum and the secondary antibody diluent are PBST solutions, and the confining liquid is a 2% BSA solution; a positive and negative critical value is 0.25. Clinical serum detection shows that the variation coefficients of in-batch and inter-batch detection results are both less than 10%; the method has good specificity; comparedwith a cELISA kit produced by the ID-Vet company for 306 parts of serum, the coincidence rate of the cELISA kit and the 306 parts of serum is 92.75%. The PPRV H protein antibody iELISA detection method can be used for clinical peste des petits ruminants antibody detection, and the operation is more time-saving, labor-saving, simple, convenient and rapid than cELISA.

Description

technical field [0001] The invention relates to the technical field of animal epidemic detection, in particular to an iELISA detection method and application of Peste des Petits Ruminants virus H protein antibody. Background technique [0002] Peste des petits ruminants (PPR) is caused by Peste des petits ruminants (PPR virus, PPRV), a member of the family Paramyxovirus of the genus Morbillivirus (Gibbs et al., 1979). A highly lethal viral disease, which is mainly transmitted through the respiratory system through direct contact, and can also be transmitted through semen, embryos, and milk. It is a highly contagious infectious disease. The main host of PPRV is small ruminants such as goats and sheep, and it can also infect large ruminants such as camels, cattle and buffaloes (Balamurugan et al., 2014; Maillard et al., 2008; Zakian et al., 2016), The clinical symptoms after infection are mainly manifested as loss of appetite, fever, increased mouth and nose secretions, and f...

AI Technical Summary

Problems solved by technology

The gene sequence encoding the N protein is located at the 3′ end of the PPRV gene sequence, which is the most abundant and most immunogenic protein in the virus. The PPRV diagnostic technology based on the N protein has also been widely developed and applied, but the protein does not induce produce immune protection

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