Direct reverse transcriptase PCR to determine virulence potential of influenza A viruses in birds.
A reverse transcriptase PCR (RT-PCR) was used for rapid determination of the hemagglutinin (HA) cleavage site sequence, a marker for the virulence potential of avian influenza viruses. When applied to specimens from chickens experimentally infected with either a virulent or an avirulent virus, RT-PCR uniformly detected the HA gene, even in specimens that were negative for virus by standard testing in eggs. This technique, combined with sequencing of the HA cleavage site, offers a rapid and sensitive way to assess the virulence potential of avian influenza viruses. Early detection of field isolates with virulence-associated structural motifs at the HA cleavage site would allow better control of influenza among large poultry populations.
Horimoto
T
T
Department of Virology and Molecular Biology, St. Jude Children's Research Hospital, Memphis, Tennessee 38101, USA.
Kawaoka
Y
Y
D000818:Animals; D001483:Base Sequence; D001717:Birds / Q000821:virology*; D002642:Chick Embryo; D002645:Chickens; D004279:DNA, Viral / Q000032:analysis; D006389:Hemagglutinins, Viral / Q000145:classification / Q000235:genetics; D009980:Influenza A virus / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity*; D005585:Influenza in Birds / Q000821:virology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D014158:Transcription, Genetic; D014774:Virulence
D004279:DNA, Viral; D006389:Hemagglutinins, Viral