Abstract: For efficient monitoring of Highly Pathogenic Avian Influenza Viruses (HPAIVs) in countries where various subtype H5 HPAIVs co-circulate in avian populations, a Simultaneous Differential Detection Method for major and minor subtypes of HPAIVs is needed. In this study, we developed a single-step Real-time Reverse Transcription-Polymerase Chain Reaction (sRRT-PCR) assay for the simultaneous detection of HPAIVM, H5, N1 and N8 genes. The M gene primer set in the assay detected all subtypes of AIV including representative H5N1 and H5N8 strains from Korea as reported in earlier studies and the H5, N1 and N8 gene primer sets were able to specifically detect H5, N1 and N8 subtypes of HPAIV, respectively. The detection limits for the M, H5, and N1 or N8 genes were 0.6, 0.7 and 0.9 EID50 for H5N1 HPAIV and 1.0, 0.8 and 1.3 EID50 for H5N8 HPAIV, respectively. The assay was sufficiently sensitive for monitoring and surveillance of HPAIVs and can be used in the differential diagnosis of H5N1, H5N8 and other subtypes of AIV in countries where subtype H5 HPAIV outbreaks are occurring.
Eun-Mi Kim and Choi-Kyu Park, 2015. Single-Step Real-Time Reverse Transcription-Polymerase Chain Reaction for Simultaneous Detection of H5N1 and H5N8 Highly Pathogenic Avian Influenza Viruses. Journal of Animal and Veterinary Advances, 14: 161-166.