Receptor Binding and Antigenic Site Analysis of Hemagglutinin Gene
Fragments of Avian Influenza Virus Serotype H5N1 Isolated from
Indonesia
Michael Haryadi Wibowo*,
Dito Anggoro, Surya Amanu, AETH Wahyuni, Tri Untari, Sidna Artanto
and Widya Asmara
Department of
Microbiology, Faculty of Veterinary Medicine, Gadjah Mada
University.Jl. Fauna 2,
Karangmalang, Yogyakarta 55281, Indonesia
*Corresponding author:
mhwibowo@ugm.ac.id; bowomikro@yahoo.com
Abstract
We reported a
retrospective study on hemagglutinin (HA) gene fragments of Avian Influenza (AI)
viruses recovered between 2010 to 2012, using reverse transcriptase polymerase
chain reaction (RT-PCR) followed by sequencing. The results provide information
about the receptor binding sites (RBS) and antigenic sites character of HA gene
of AI viruses in Indonesia. Viral RNA was extracted from allantoic fluid of
specific pathogen free (SPF) of chicken embryonated eggs inoculated by AI
suspected samples. Amplification was performed by using H5 specific primers to
produce amplification target of 544 bp. The resulting sequences were analyzed
with MEGA-5 consisting of multiple alignment, deductive amino acid prediction,
and phylogenetic tree analysis. The results showed that out of the 12 samples
amplified using RT-PCR technique, only 7 were detected to be avian influenza
serotype H5 viruses. Sequence analysis of AIV H5 positive samples, showed a
binding preference towards avian type receptors. Antigenic site analysis is
consistent with the previous report, however, the antigenic site B at position
189 showed that the residue had undergone mutation from arginine to methionine.
Phylogenetic tree analysis showed that these viruses were clustered into clade
2.1.3. Our report supports the importance of the previous study of RBS and
antigenic properties of HPAI H5N1 in Indonesia.