Development and evaluation of an indirect ELISA based on recombinant structural protein σC to detect antibodies against avian orthoreovirus.

The avian orthoreovirus (ARV) causes large economic losses to poultry industry around the whole world. ARV viral proteins can be classified into three sizes: λ (large), μ (medium) and σ (small). σC, one of the capsid protein of ARV, contains various specific neutralizing epitopes and can induce robust immune responses in infected chickens. An indirect enzyme-linked immunosorbent assay (I-ELISA) was established using the recombinant σC protein. The optimal dilutions of antigen, serum and conjugate goat anti-chicken IgY conjugate were 2.5 μg/ml, 1:80 and 1:4000, respectively. The optimal antigen coating condition was 4℃ for overnight. Hyperimmune serum of high-yielding hens of different age produced stronger reaction against σC. Chicken serum samples were randomly collected and total coincidence rate between commercial kit and σC-ARV-ELISA kit was 97.8%. Importantly, those discrepant serums were further verified using Western blot assay and three of them were determined to be positive. Thus, the method based on σC protein has higher sensitivity compared with the commercial kit and provide a rapid and simple method for serodiagnosis of ARV, contributing to convenient epidemiological monitoring of ARV.
Competing Interests: Declaration of Competing Interest The authors declare no commercial of financial conflict of interest.
(Copyright © 2021. Published by Elsevier B.V.)