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A novel monoclonal antibodies-based sandwich ELISA for detection of serotype 4 fowl adenovirus

Shao, Hongxia, Wang, Ping, Wang, Weikang, Zhang, Jianjun, Li, Tuofan, Liang, Guangchen, Gao, Wei, Qin, Aijian, Ye, Jianqiang
Avian pathology 2019 v.48 no.3 pp. 204-208
antigens, chickens, detection limit, enzyme-linked immunosorbent assay, financial economics, flocks, monoclonal antibodies, poultry industry, serotypes, viruses
As a major causative agent for hepatitis-hydropericardium syndrome (HPS) in chickens, serotype 4 fowl adenovirus (FAdV-4) has caused huge economic losses in the poultry industry globally. However, there is no commercial diagnostic test for FAdV-4 antigens. To generate a rapid approach for specific detection of FAdV-4, a monoclonal antibodies (mAbs)-based sandwich ELISA was developed. In this ELISA, a purified mAb 4A3 and a HRP-labelled mAb 3C2 specific to the fiber-2 of FAdV-4 were used as the capture antibody and detection antibody respectively. Specificity assay revealed the ELISA only reacted with FAdV-4, not with other avian viruses tested. Sensitivity assay showed the limit of detection of the ELISA was 1000 TCID₅₀/ml and 12.5 ng/ml for the FAdV-4 and the purified GST-Fiber2 protein respectively. Moreover, the ELISA could be efficiently applied in detecting the FAdV-4 in tissue samples from a clinically-diseased chicken flock. All these data demonstrated that the ELISA developed here provides a promising tool for rapid and efficient diagnosis of clinical infection with FAdV-4.