Type-specific identification of influenza viruses A, B and C by the polymerase chain reaction.
The aim of this study was to develop a polymerase chain reaction for specific detection of influenza A, B, and C RNA genomes. Three primer sets were selected from conserved regions of the genome coding for the non-structural proteins and were tested on 61 influenza A (22 H1N1, 9 H2N2, and 30 H3N2), 11 influenza B, and three influenza C isolates. Specific amplified products were obtained with all these strains after electrophoresis on a 2% agarose gel. The specificity of the reaction was increased by hybridization with oligonucleotide probes. When nucleic acids from a variety of micro-organisms from the respiratory tract were subjected to the PCR with these primers, no specific amplified products were generated. The sensitivity of the technique was found to be at the subpicogram level. The RNA-PCR was applied to 21 clinical specimens from patients with a culture/IF proven influenza infection. Six influenza A positive patients and 13 influenza B positive patients could be confirmed in the RNA-PCR. In two cases, influenza B positive IF specimens were found negative by the PCR. No virus could be isolated on eggs or tissue culture from these samples. RNA-PCR is a specific and sensitive technique for the detection of influenza virus genomes.
Claas
E C
EC
Department of Virology, Erasmus University, Rotterdam, Netherlands.
Sprenger
M J
MJ
Kleter
G E
GE
van Beek
R
R
Quint
W G
WG
Masurel
N
N
D001483:Base Sequence; D015342:DNA Probes; D004279:DNA, Viral / Q000235:genetics; D005069:Evaluation Studies as Topic; D006801:Humans; D008969:Molecular Sequence Data; D009975:Orthomyxoviridae / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012367:RNA, Viral / Q000235:genetics; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*
D015342:DNA Probes; D004279:DNA, Viral; D012367:RNA, Viral