Development and Optimization of Multiplex PCR for the Identification of Different Strains of Avian Influenza Viruses in Pakistan

†M. S. Saleem, ‡Z. Mahel, *A. Mushtaq, ‡A. Aslam and A. Mushtaq

†Department of Microbiology, ‡Department of Pathology, University of Veterinary and Animal Sciences, Lahore, Pakistan, *Veterinary Research Institute Lahore, Pakistan.

ABSTRACT

The present study was conducted to develop and optimize a multiplex reverse transcriptase-polymerase chain reaction (mRT-PCR) assay for the rapid detection of type A Avian influenza (AI) viruses. The assay simultaneously differentiates avian H5, H7 and H9 hemagglutinin subtypes. Specific primers were used in this assay for H5, H7 and H9 heamagglutinin subtypes. The mRT-PCR DNA products were visualized by gel electrophoresis. The fragments observed were of 362bp, 699bp and 287bp for H5, H7 and H9 hemagglutinin subtypes, respectively. The multiplex PCR is rapid and cost effective compared to other available assays for AI in Pakistan.

Key words: Avian Influenza, Multiplex PCR, Virus, Pakistan.