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 us...

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Bibliographic Details
Published inMicrobiology and immunology Vol. 54; no. 3; p. 129
Main Authors Inoue, Emi, Wang, Xiaofeng, Osawa, Yoshiaki, Okazaki, Katsunori
Format Journal Article
LanguageEnglish
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
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Summary: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.
ISSN:0385-5600
DOI:10.1111/j.1348-0421.2009.00193.x