Full genomic amplification and subtyping of influenza A virus using a single set of universal primers.
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.
Inoue
Emi
E
Laboratory of Microbiology and Immunology, Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido, Ishikari-tobetsu, Hokkaido, Japan.
Wang
Xiaofeng
X
Osawa
Yoshiaki
Y
Okazaki
Katsunori
K
D000818:Animals; D002642:Chick Embryo; D017931:DNA Primers / Q000235:genetics*; D016679:Genome, Viral; D006389:Hemagglutinins, Viral / Q000235:genetics; D009980:Influenza A virus / Q000145:classification / Q000235:genetics*; D009439:Neuraminidase / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014764:Viral Proteins / Q000235:genetics
D017931:DNA Primers; D006389:Hemagglutinins, Viral; D014764:Viral Proteins; C487630:NA protein, influenza A virus; D009439:Neuraminidase