Abstract
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.
Key points
• B-cell epitopes of GAstV capsid protein were predicted and expressed as immunogen
• A core antigenic advantage domain-based ELISA was established to detect GAstV-specific antibody
• The established ELISA will contribute to inspection and elimination of infected breeding geese and provide a useful tool for large scale serological testing of GAstV in geese
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All the data generated or analyzed during this study are included in this published article and its supplementary information files.
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Funding
This work was supported by grants from the National Natural Science Foundation of China (grant no. 32002259), Natural Science Foundation of Henan Province (grant no. 202300410198), Youth Science Foundations of Henan University of Animal Husbandry and Economy (2019HNUAHEDF008), Veterinary Discipline Key Construction Project of Henan University of Animal Husbandry and Economy (MXK2016102) and Scientific Research and Innovation Team in Henan University of Animal Husbandry and Economy (2018KYTD18).
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ZW, XZ, and JZ conceived and designed the study. HC, SG, XZ, MS, and ZS performed the experiments. QJ, ZP, LZ, HQ, CB, XY, ZW, and JZ analyzed the experiment data. ZW, XZ, and JZ wrote and revised the manuscript. All the authors reviewed the manuscript and approved the final version.
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Not applicable. The experiments in this study did not involve animal experiments and no ethical policy was approved.
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Wang, Z., Chen, H., Gao, S. et al. Core antigenic advantage domain–based ELISA to detect antibody against novel goose astrovirus in breeding geese. Appl Microbiol Biotechnol 106, 2053–2062 (2022). https://doi.org/10.1007/s00253-022-11852-y
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DOI: https://doi.org/10.1007/s00253-022-11852-y