Full genomic amplification and subtyping of influenza A virus using a single set of universal primers

• Published in: Microbiology and immunology Vol. 54; no. 3; p. 129
• Main Authors: Inoue, Emi, Wang, Xiaofeng, Osawa, Yoshiaki, Okazaki, Katsunori
• Format: Journal Article
• Language: English
• Published: Australia 01.03.2010
• Subjects: Animals; Chick Embryo; DNA Primers - genetics; Genome, Viral; Hemagglutinins, Viral - genetics; Influenza A virus - classification; Influenza A virus - genetics; Neuraminidase - genetics; Reverse Transcriptase Polymerase Chain Reaction - methods; Viral Proteins - genetics
• DOI: 10.1111/j.1348-0421.2009.00193.x
• ISSN: 0385-5600

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.