Genetic subtyping of influenza A viruses using RT-PCR with a single set of primers based on conserved sequences within the HA2 coding region.
Influenza A viruses are subtyped conventionally according to the antigenic characteristics of the external glycoproteins, haemagglutinin (HA) and neuraminidase (NA). To date 15 HA and 9 NA subtypes have been described. There is a need to develop fast, accurate and reliable methods to identify influenza virus subtypes, which may be associated with disease outbreaks. An RT-PCR is described using a single primer pair based on a conserved region of the HA2 gene that can detect all 15 HA influenza A subtypes. The assay was validated initially using a panel of 12 known standard prototype strains of influenza virus representing 6 HA subtypes and subsequently in a blind study using a panel of 30 strains. Selected viruses represented all known HA subtypes derived from avian, swine and human hosts separated both geographically and with time Sequence analysis of RT-PCR product showed complete correlation with results obtained using conventional serological methods. It is concluded that this RT-PCR is a reliable, robust and reproducible tool for the rapid identification of a wide range of all the HA subtypes of influenza A viruses.
Virology Department, Veterinary Laboratories Agency, New Haw, Addlestone, Surrey, UK.
L P
LP
Phipps
S C
SC
Essen
I H
IH
Brown
D017931:DNA Primers; D005838:Genotype; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics*; D009980:Influenza A virus / Q000145:classification* / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; D019267:Hemagglutinin Glycoproteins, Influenza Virus