Core antigenic advantage domain-based ELISA to detect antibody against novel goose astrovirus in breeding geese
文献类型： 外文期刊
作者： Wang, Zeng 1 ; Chen, Huayuan 1 ; Gao, Shenyan 1 ; Song, Mingzhen 1 ; Shi, Zicong 1 ; Peng, Zhifeng 2 ; Jin, Qianyue 3 ; Zhao, Li 2 ; Qiao, Hongxing 2 ; Bian, Chuanzhou 2 ; Yang, Xia 1 ; Zhang, Xiaozhan 2 ; Zhao, Jun 1 ;
作者机构： 1.Henan Agr Univ, Coll Vet Med, Zhengzhou, Peoples R China
2.Henan Univ Anim Husb & Econ, Coll Vet Med, Zhengzhou, Peoples R China
3.Henan Acad Agr Sci, Henan Prov Key Lab Anim Immunol, Zhengzhou, Peoples R China
关键词： Goose astrovirus; Capsid protein; Antigenic advantage domain; Indirect ELISA; Serological investigation
期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：5.56； 五年影响因子：5.365 ）
ISSN： 0175-7598
年卷期： 2022 年 106 卷 5-6 期
页码：
收录情况： SCI
摘要： Goose astrovirus (GAstV), the major causative agent of visceral and joint gout in goslings, is a novel pathogen greatly threatening waterfowl industry. Importantly, the horizontal and vertical transmissibility of GAstV posed a great challenge for disease prevention and control. Given the absence of commercial vaccine, restricting vertical transmission and protecting susceptible goslings must be a priority. Although many detection methods have been established, there is no serological method to detect GAstV-specific antibody, greatly limiting inspection and elimination of infected breeding bird. In this study, the B-cell epitopes of GAstV capsid protein were predicted, and its core antigenic advantage domain (shCAP) was expressed and purified. After authenticating the antigenicity, the recombinant shCAP protein was taken as the coating antigen, and an easily accessible indirect enzyme-linked immunosorbent assay (ELISA) was established to detect GAstV-specific antibody. The working conditions, including antigen concentration, serum dilution and incubation time, blocking buffer concentration, and color developing time, were gradually optimized by checkerboard titration. The cut-off OD450 value of the indirect ELISA for positive sample was 0.379, and the analytical sensitivity was 1:800. There was no cross-reaction with sera against goose parvovirus (GPV), Tembusu virus (TUMV), H5 and H7 subtype avian influenza virus (AIV H5 + H7), and Newcastle disease virus (NDV). The assay was further applied to examine 73 breeding goose serum samples and shared excellent agreement of 93.5% (68/73) with western blot, which also suggested that GAstV is circulating in the goose population in China. In conclusion, the developed indirect ELISA is simple, specific, and sensitive, which will be greatly useful to screen GAstV infection and block vertical transmission.
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