Development and evaluation of an indirect ELISA based on recombinant structural protein VP2 to detect antibodies against duck hepatitis A virus

Yalan Lai; Ling He; Anchun Cheng; Mingshu Wang; Xumin Ou; XingJian Wen; Sai Mao; Di Sun; Renyong Jia; Qiao Yang; Ying Wu; Dekang Zhu; Shun Chen; Mafeng Liu; Shaqiu Zhang; Xin-Xin Zhao; Juan Huang; Qun Gao; Yunya Liu; Yanling Yu; Ling Zhang; Bin Tian; Leichang Pan; Mujeeb Ur Rehman; Xiaoyue Chen

doi:10.1016/j.jviromet.2020.113903

Development and evaluation of an indirect ELISA based on recombinant structural protein VP2 to detect antibodies against duck hepatitis A virus

J Virol Methods. 2020 May 30:282:113903. doi: 10.1016/j.jviromet.2020.113903. Online ahead of print.

Authors

Yalan Lai¹, Ling He¹, Anchun Cheng¹, Mingshu Wang², Xumin Ou¹, XingJian Wen¹, Sai Mao¹, Di Sun¹, Renyong Jia¹, Qiao Yang¹, Ying Wu¹, Dekang Zhu³, Shun Chen¹, Mafeng Liu¹, Shaqiu Zhang¹, Xin-Xin Zhao¹, Juan Huang¹, Qun Gao¹, Yunya Liu¹, Yanling Yu¹, Ling Zhang¹, Bin Tian⁴, Leichang Pan⁴, Mujeeb Ur Rehman⁴, Xiaoyue Chen¹

Affiliations

¹ Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu City, Sichuan 611130, PR China; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Wenjiang, Chengdu City, Sichuan 611130, PR China; Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu City, Sichuan 611130, PR China.
² Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu City, Sichuan 611130, PR China; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Wenjiang, Chengdu City, Sichuan 611130, PR China; Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu City, Sichuan 611130, PR China. Electronic address: mshwang@163.com.
³ Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Wenjiang, Chengdu City, Sichuan 611130, PR China; Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu City, Sichuan 611130, PR China.
⁴ Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu City, Sichuan 611130, PR China; Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu City, Sichuan 611130, PR China.

PMID: 32485472
DOI: 10.1016/j.jviromet.2020.113903

Abstract

An indirect enzyme-linked immunosorbent assay (I-ELISA) based on the VP2 protein of duck hepatitis A virus type 3 (DHAV-3) was established in this study. The optimal dilutions of antigen, serum and goat anti-duck IgG conjugate were 1:1600 (2.23 μg/mL), 1:160 and 1:2000, respectively. The optimal blocking buffer was 1% skim milk. The cut-off value for the method was 0.25, and the analytical sensitivity of the method was 1:5120. The results of specific evaluation showed that except for DHAV-1, DHAV-3 antisera did not cross-react with any other common duck-sensitive pathogens, indicating that this method can be used to detect DHAV-3 and DHAV-1 antibodies. The coefficients of variation (CVs) were lower than 10 %. The coincidence rate between the VP2-DHAV-3-ELISA and the neutralization test was 93.3 %. In summary, the I-ELISA method based on VP2 protein has high sensitivity, specificity, and coincidence rate compared with the neutralization test and has advantages in serum monitoring. The I-ELISA method based on VP2 protein provides a simple and rapid method for the detection of anti-DHAV antibodies and the epidemiological monitoring of DHAV.

Keywords: Antibody detection; Duck hepatitis A virus type 3; Indirect ELISA (I-ELISA); VP2 protein.