title: Evaluation of a conserved HA274-288 epitope to detect antibodies to highly pathogenic avian influenza virus H5N1 in Indonesian commercial poultry.
abstract: A peptide enzyme linked immunosorbent assay (ELISA) based on an epitope in the haemagglutinin (HA) of avian influenza virus H5N1, amino acid positions 274-288 (HA274-288) was evaluated for detection of H5N1-specific antibodies. An optimized ELISA based on the tetrameric form of the HA274-288 epitope designated MP15 gave low background with non-immune chicken sera and detected vaccinated and infected birds. The HA274-288 epitope was highly conserved in Indonesian H5N1 strains and antibody responses were detected in the majority of the vaccinated chickens regardless of the H5N1 strain used for vaccination. The HA274-288 epitope was also conserved in the majority of H5N1 strains from the neighbouring Asian region, and other H5 subtypes potentially allowing for a wider use of the MP15 ELISA in H5N1 vaccinated and infected flocks. The MP15 ELISA results correlated significantly with haemagglutination inhibition (HI) test results and test sensitivity and specificity were 87% and 92%, respectively. The MP15 ELISA titres were significantly higher than the HI titres in all immune sera allowing for sera to be tested at a single dilution of 1:400 which is of advantage in routine surveillance. The study indicated that the MP15 ELISA is potentially useful for serological detection of H5N1 vaccinated or infected poultry and to have some advantages over the standard HI test for routine monitoring of flocks' immunity after vaccination.
authors.affiliation: a Faculty of Veterinary and Agricultural Sciences , University of Melbourne , Melbourne , Australia.
authors.forename: Nadeeka K
authors.initials: NK
authors.lastname: Wawegama
authors.affiliation: b Indonesian Research Centre for Veterinary Science , Bogor , Indonesia.
authors.forename: Simson
authors.initials: S
authors.lastname: Tarigan
authors.affiliation: b Indonesian Research Centre for Veterinary Science , Bogor , Indonesia.
authors.forename: Risa
authors.initials: R
authors.lastname: Indriani
authors.affiliation: c CSIRO, Australian Animal Health Laboratory , Geelong , Australia.
authors.forename: Paul
authors.initials: P
authors.lastname: Selleck
authors.affiliation: b Indonesian Research Centre for Veterinary Science , Bogor , Indonesia.
authors.forename: Rm Abdul
authors.initials: RA
authors.lastname: Adjid
authors.affiliation: b Indonesian Research Centre for Veterinary Science , Bogor , Indonesia.
authors.forename: Tati
authors.initials: T
authors.lastname: Syafriati
authors.affiliation: b Indonesian Research Centre for Veterinary Science , Bogor , Indonesia.
authors.lastname: Hardiman
authors.affiliation: c CSIRO, Australian Animal Health Laboratory , Geelong , Australia.
authors.forename: Peter A
authors.initials: PA
authors.lastname: Durr
authors.affiliation: a Faculty of Veterinary and Agricultural Sciences , University of Melbourne , Melbourne , Australia.
authors.forename: Jagoda
authors.initials: J
authors.lastname: Ignjatovic
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D002645:Chickens / Q000821:virology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D000939:Epitopes / Q000276:immunology*; D006385:Hemagglutination Inhibition Tests / Q000662:veterinary; D007214:Indonesia / Q000453:epidemiology; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology*; D005585:Influenza in Birds / Q000517:prevention & control* / Q000821:virology; D011200:Poultry; D011201:Poultry Diseases / Q000517:prevention & control* / Q000821:virology; D012680:Sensitivity and Specificity; D014611:Vaccination / Q000662:veterinary
chemical_list: D000914:Antibodies, Viral; D000939:Epitopes
keywords: H5N1 in Indonesia; H5N1 vaccinated commercial poultry; Highly pathogenic avian influenza; peptide ELISA; serological detection of H5N1