Tissue culture propagation of highly pathogenic avian influenza H5N1 virus

Document Type : Original Article

Authors

Central Laboratory for Evaluation of Veterinary Biologics, Abbasia, Cairo.

Abstract

   Avian influenza virus usually refers to influenza A viruses found chiefly in birds, but infections can occur in humans. The risk is generally low to most people, because the viruses do not usually infect humans. However, confirmed cases of human infection have been reported since 1997 by H5N1 subtype. In the current study, a highly pathogenic avian influenza subtype HSNI was isolated and confirmed by PCR and sequencing. Sequence analysis revealed a high degree of identity with other published sequences of H5N1 isolates from Egypt. The multiple basic amino acid sequence of the isolated virus was similar to the highly pathogenic strains of H5N1 (G-E/R-R- R/K-K-R.) and to other viruses isolated in Egypt in the previous years specially in the presence of arginine amino acid in the cleavage site motive of the hemagglutinin protein. 
The isolated virus which designed as A/chicken/1000CLEVB/2010 was propagated on chicken embryo fibroblast tissue culture. The virus titer was rapidly increasing from 4.8 log10 /ml in the 1st passage to 8.5 log10/ml by the 3rd one. The HA properties of the virus was regained by treatment with 0.5µg of trypsin/10 min which open the gates for the production of inactivated tissue culture propagated vaccine.

Main Subjects