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Avian Pathol. 2019 Nov 18:1-27. doi: 10.1080/03079457.2019.1694635. [Epub ahead of print] Analysis of Antibody Response to an Epitope in the Haemagglutinin Subunit 2 of Avian Influenza Virus H5N1 for Differentiation of Infected and Vaccinated Chickens.
Putri K1, Wibowo MH2, Tarigan S3, Wawegama N1, Ignjatovic J1, Noormohammadi AH1.
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1 Asia Pacific Centre for Animal Health, Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Werribee, Victoria, Australia 3030. 2 Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia 55281. 3 Indonesian Research Centre for Veterinary Science (IRCVS), Bogor, Jawa Barat, Indonesia 116114.
Abstract
The H5N1 subtype of highly pathogenic avian influenza virus (HPAIV) has been circulating in poultry in Indonesia since 2003 and vaccination has been used as a strategy to eradicate the disease. However, monitoring of vaccinated poultry flocks for H5N1 infection by serological means has been difficult, as vaccine antibodies are not readily distinguishable from those induced by field viruses. Therefore a test that differentiates infected and vaccinated animals (DIVA) would be essential. Currently, no simple and specific DIVA test is available for screening of a large number of vaccinated chickens. Several epitopes on E29 domain of the haemagglutinin H5N1 subunit 2 (HA2), have been recently examined for their antigenicity and potential as possible marker for DIVA in chicken. In this study, the potential of E29 as antigen for DIVA was evaluated in details. Three different forms of full-length E29 peptide, a truncated E29 peptide (E15) and a recombinant E29, were compared for their ability to detect anti E29 antibodies. Preliminary ELISA experiments using mono-specific chicken and rabbit E29 sera, and a mouse monoclonal antibody revealed that the linear E29 peptide was the most antigenic. Further examination of the E29 antigenicity in ELISA using several experimentally infected or vaccinated chicken sera revealed that the full length E29 peptide had the greatest discrimination power between infected and vaccinated chicken sera while provided the least non-specific reaction. This study demonstrates the usefulness of the HPAI H5N1 HA2 E29 epitope as a DIVA antigen in HPAI H5N1 vaccinated and infected chickens.Research Highlight E29 (HA2 position 488-516) epitope is antigenic in chickenAntibodies to E29 are elicited following live H5N1 virus infection in chickenE29 epitope is a potential DIVA antigen for use in ELISA.
KEYWORDS:
Avian Influenza; DIVA; ELISA; H5N1; HA2; poultry
PMID: 31738584 DOI: 10.1080/03079457.2019.1694635