A rapid method for the analysis of influenza virus genes: application to the reassortment of equine influenza virus genes.
We describe a rapid method for genetic characterisation of influenza virus genes using reverse transcription and amplification by polymerase chain reaction (RT/PCR) of all virus segments simultaneously (multiplex RT/PCR) using primers based on the conserved terminal sequences. The product has been shown to be suitable for determination of partial nucleotide sequences which can be used to search nucleotide sequence databases and rapidly map the genetic origin of each segment. We illustrate the use of the method by analysing genetic reassortment in H7N7 equine influenza viruses.
Adeyefa
C A
CA
AFRC Institute for Animal Health, Pirbright Laboratory, Woking, Surrey, UK.
Quayle
K
K
McCauley
J W
JW
D001483:Base Sequence; D017124:Conserved Sequence; D017931:DNA Primers; D016208:Databases, Factual; D005814:Genes, Viral; D009980:Influenza A virus / Q000235:genetics*; D008969:Molecular Sequence Data; D009975:Orthomyxoviridae / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics*; D012194:RNA-Directed DNA Polymerase; D016865:Reassortant Viruses / Q000235:genetics*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
D017931:DNA Primers; D012367:RNA, Viral; D012194:RNA-Directed DNA Polymerase