DEVELOPMENT OF MULTIPLEX- RT-PCR FOR RAPID DETECTION AND SUBTYPING OF AVIAN INFLUENZA VIRUSES TYPE A

A multiplex reverse transcription polymerase chain reaction (RT-PCR) was developed and optimized to simultaneously detect 3 subtypes (H5, H7 and H9) of avian influenza virus type A. The developed method was specific to amplify only the HA gene of H5, H7 and H9 of Influenza type A and no amplification with other subtypes of AIV and other avian infectious viruses such as Newcastle disease virus, infectious bronchitis virus, infectious bursal disease virus was observed. The multiplex RT-PCR assay developed in this study was as sensitive as conventional RT-PCR and virus isolation. Therefore this method is reliable, rapid, sensitive and specific and could be used as a tool for detecting and subtyping of AIV, in the control program of AIV.