Archives of Virology

, Volume 162, Issue 2, pp 487–494 | Cite as

Antigenic characterization of H5N1 highly pathogenic avian influenza viruses isolated from poultry in India, 2006-2015

  • Sudipta Bhat
  • Shanmugasundaram Nagarajan
  • Manoj Kumar
  • Harshad V. Murugkar
  • Semmannan Kalaiyarasu
  • Govindarajulu Venkatesh
  • Chakradhar Tosh
Original Article


Highly pathogenic avian influenza (HPAI) is a major health concern worldwide. In this study, we focused on antigenic analysis of HPAI H5N1 viruses isolated from poultry in India between 2006 and 2015 comprising 25 isolates from four phylogenetic clades 2.2 (1 isolate), (1 isolate), (17 isolates) and (6 isolates). Seven H5N1 isolates from all four clades were selected for production of chicken antiserum, and antigenic analysis was carried out by hemagglutination inhibition (HI) assay. HI data indicated antigenic divergence (6-21 fold reduction in cross-reactivity) between the two recently emerged clades and These two clades are highly divergent (21-128 fold reduction in HI titre) from the earlier clades 2.2 / isolated in India. However, a maximum of 2-fold and 4-fold reduction in cross-reactivity was observed within the isolates of homologous clades and, respectively. The molecular basis of inter-clade antigenic divergence was examined in the haemagglutinin (HA) antigenic sites of the H5N1 virus. Amino acid changes at 8 HA antigenic sites were observed between clades and, whereas 20-23 substitutions were observed between clades and 2.2/ Therefore, a systematic analysis of antigenic drift of the contemporary field isolates is a pre-requisite for determining the suitable strain(s) for vaccine candidature.


Influenza Avian Influenza H5N1 Virus Avian Influenza Virus Haemagglutination Inhibition 



We are thankful to the ICAR, New Delhi and Director, ICAR-NIHSAD, Bhopal for providing necessary facilities to carry out this work. Funding by DADF, MoA, GOI through CDDL grant is acknowledged. S. Bhat acknowledges Director, ICAR-Indian Veterinary Research Institute for financial support.

Compliance with ethical standards


This study was supported by CDDL Grant of DADF, Ministry of Agriculture and Farmers Welfare, Government of India. S. Bhat received junior research fellowship from ICAR-Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh.

Conflicts of interest

The authors declare that they have no conflict of interest.

Ethical Standard

The animal experiment was carried out as per the guidelines of CPCSEA, Government of India with approval from Institutional Animal Ethics Committee (IAEC Approval No. 54/IAEC/HSADL/11).


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Copyright information

© Springer-Verlag Wien 2016

Authors and Affiliations

  • Sudipta Bhat
    • 1
    • 2
  • Shanmugasundaram Nagarajan
    • 1
  • Manoj Kumar
    • 1
  • Harshad V. Murugkar
    • 1
  • Semmannan Kalaiyarasu
    • 1
  • Govindarajulu Venkatesh
    • 1
  • Chakradhar Tosh
    • 1
  1. 1.ICAR-National Institute of High Security Animal DiseasesBhopalIndia
  2. 2.ICAR-Indian Veterinary Research InstituteBareillyIndia

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