An enzyme-linked immunosorbent assay for detection of avian influenza virus subtypes H5 and H7 antibodies

Trine H Jensen,Kurt J Handberg,Gitte Ajjouri,Martine Cherbonnel,Véronique Jestin,Vivien J Coward,Peter Lind,Marek J Slomka,Poul H Jørgensen

doi:10.1186/1751-0147-55-84

Abstract

BackgroundAvian influenza virus (AIV) subtypes H5 and H7 attracts particular attention because of the risk of their potential pathogenicity in poultry. The haemagglutination inhibition (HI) test is widely used as subtype specific test for serological diagnostics despite the laborious nature of this method. However, enzyme-linked immunosorbent assays (ELISAs) are being explored as an alternative test method.H5 and H7 specific monoclonal antibodies were experimentally raised and used in the development of inhibition ELISAs for detection of serological response specifically directed against AIV subtypes H5 and H7. The ELISAs were evaluated with polyclonal chicken anti-AIV antibodies against AIV subtypes: H1N2, H5N2, H5N7, H7N1, H7N7, H9N9, H10N4 and H16N3.ResultsBoth the H5 and H7 ELISA proved to have a high sensitivity and specificity and the ELISAs detected H5 and H7 antibodies earlier during experimental infection than the HI test did. The reproducibility of the ELISA’s performed at different times was high with Pearson correlation coefficients of 0.96-0.98.ConclusionsThe ELISAs are a potential alternative to the HI test for screening of large amounts of avian sera, although only experimental sera were tested in this study.

Highlights

Avian influenza virus (AIV) subtypes H5 and H7 attracts particular attention because of the risk of their potential pathogenicity in poultry
Specificity of the H5 and H7 monoclonal antibody (mAb) The H5 and H7 mAbs were specific as they recognised only H5 and H7 subtype AIV strains shown by immunocytochemistry and direct enzyme-linked immunosorbent assay (ELISA) coated with various AIV strains (Table 2)
Specificity and sensitivity of the H5 inhibition ELISA For studies of specificity heterologous sera from chickens immunised with PMV8, H16N3, H1N2, H9N9, H10N4 and H7N1 were chosen

Summary

Introduction

Avian influenza virus (AIV) subtypes H5 and H7 attracts particular attention because of the risk of their potential pathogenicity in poultry. The zoonotic potential of H5 and H7 infections [3,4] and the severe impact of HPAI infections for the poultry industry [5] emphasise the need for sensitive and effective diagnostic methods and surveillances to early detect low (AGID) test [16,17,18,19,20]. These kits had higher sensitivity compared to the AGID when testing duck and wild bird sera [16,19,21]. Two promising studies of H5 ELISA using H5 mAb was recently described for testing of chickens, turkeys and ducks [25,28]

Methods

Results

Discussion

Conclusion