Full genomic amplification and subtyping of influenza A virus using a single set of universal primers |
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Authors: | Emi Inoue Xiaofeng Wang Yoshiaki Osawa Katsunori Okazaki |
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Institution: | 1. Laboratory of Microbiology and Immunology, Faculty of Pharmaceutical Sciences;2. Department of Microbiology, Medical College of Qingdao University, Quingdao 266021, China;3. Division of Medicinal Science, The Research Institute of Personalized Health Sciences, Health Sciences University of Hokkaido, Ishikari‐Tobetsu, Hokkaido 061‐0293, Japan |
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Abstract: | Influenza A virus has eight‐segmented RNA molecules as a genome and, among all strains of the virus, both ends of each segment have 13 and 12 nucleotide sequences conserved. In the present study, a simple RT‐PCR method to amplify all eight segments of the virus and determine the HA and NA subtype using a single primer set based on the conserved terminal sequences has been established. This method is also capable of detecting subgenomic defective interfering RNA of the influenza A virus. Since the primers used here cope with each and every RNA segment of influenza A virus, this simple RT‐PCR method is valuable not only for cloning each gene of the virus, but also for identifying subtypes, including subtypes other than 16 HA and 9 NA subtypes. |
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Keywords: | genomic amplification influenza A virus single primer set subtyping |
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