Development of an indirect enzyme-linked immunosorbent assay for the detection of novel chicken orthoreovirus.

Abstract

A novel avian orthoreovirus (N-ARV) variant characterized with obvious arthritis and synovial inflammation, was isolated from Shandong, China in May 2016. It caused chicken poor growth and enormous economic losses to the poultry industry of China. However, there are few effective methods for detecting the antibody levels of N-ARV. In this study, a viral structural protein σC was expressed using the prokaryotic expression vector pET32a (+). The target protein was obtained by inducing for 6 hours at an IPTG concentration of 0.6mM. The optimal dilution of the coating antigen and serum antibody were determined to be 1000 fold and 10 fold respectively. A specificity test showed that there was no positive reactivity between N-ARV and other pathogens, and when the positive serum was diluted 100 times detection results were still checkable. The repeatability of this method was determined by the inter assay and intra assay tests with variability ranging from 4.85% to 7.93%. In conclusion, this indirect enzyme linked immunosorbent assay (ELISA) will be useful for large-scale serological surveys and monitoring antibody levels in N-ARV infection.