Serological investigation of highly pathogenic avian influenza H5N2 in ostriches (Struthio camelus)

• Published in: Avian pathology 2013-06, Vol.42 (3), p.206-214
• Main Authors: Abolnik, Celia, Fehrsen, Jeanni, Olivier, Adriaan, van Wyngaardt, Wouter, Fosgate, Geoffrey, Ellis, Charlotte
• Format: Article
• Language: English
• Subjects: Animals; antibodies; Antigens; Avian flu; avian influenza; Avian influenza virus; Birds; chicken eggs; chickens; Comparative analysis; detection limit; enzyme-linked immunosorbent assay; Enzyme-Linked Immunosorbent Assay - methods; Enzyme-Linked Immunosorbent Assay - veterinary; farms; flocks; Glycoproteins; hemagglutination; Hemagglutination Tests - veterinary; Hemagglutinin Glycoproteins, Influenza Virus - genetics; immunoglobulin Y; Influenza A virus; Influenza A Virus, H5N2 Subtype - genetics; Influenza in Birds - epidemiology; Influenza in Birds - virology; ostriches; peroxidase; Polymerase chain reaction; Sensitivity and Specificity; Seroepidemiologic Studies; South Africa; Struthio camelus; Struthioniformes; vaccination
• ISSN: 1465-3338; 0307-9457; 1465-3338
• DOI: 10.1080/03079457.2013.779637

An ostrich farm of 929 birds that tested polymerase chain reaction-positive for highly pathogenic avian influenza H5N2 in a single sample was designated for culling, despite no evidence of sero-conversion as assessed by haemagglutination inhibition (HI) tests. A month later and immediately prior to culling, all birds were bled and tested with an IDEXX avian influenza virus (AIV) nucleoprotein (NP)-specific enzyme-linked immunosorbent assay (ELISA) and a high sero-prevalence was detected. To address the question of whether the NP-specific antibodies detected indicated exposure to H5 or non-H5 subtypes (H6N2 and H1N2 strains were also circulating regionally at the time), we developed two H5-specific ELISAs, both based on a recombinant H5 HA1 antigen. The H5 indirect ELISA used a horseradish peroxidase ostrich IgY conjugate that we produced in chicken eggs. The single-chain variable fragment (scFv) competitive ELISA (H5 scFv cELISA) used a scFv derived from an H5-immune chicken scFv library. By comparing IDEXX AIV ELISA results with those of the two H5-specific ELISAs and HI tests, we determined that up to 89% of the flock had been exposed to H5N2 AIV. We also detected evidence of suspected vaccination, since 17% of sera contained antibodies against the H5 glycoprotein but not the NP protein. Comparative analytical sensitivity indicated that HI tests are likely to miss up to 35% of H5-positive samples, and thus we consider that H5/H7-specific ELISAs should replace HI tests for ostrich testing in future.