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J Virol Methods. 2004 May;117(2):103-12.

Rapid detection and simultaneous subtype differentiation of influenza A viruses by real time PCR.

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1
Infectious Diseases Laboratory, Institute of Medical and Veterinary Science, P.O. Box 14, Frome Road, South Australia, Rundle Mall, S.A. 5000, Australia.

Abstract

A real time RT-PCR, using the LightCycler, was developed and compared with rapid antigen enzyme immunoassay (AgEIA) and enhanced virus culture for rapid detection of influenza A viruses in stored and prospectively collected respiratory specimens. Specific hybridization probes were used for simultaneous detection and differentiation between H1N1 and H3N2 subtypes. The sensitivity of the RT-PCR for influenza A H1N1 was 120 copies and H3N2 350 copies of in vitro transcribed RNA. A specimen was considered positive for influenza A when it was culture positive or at least two methods yielded a positive test result. Using these criteria, with stored samples, the RT-PCR sensitivity, specificity, positive and negative predictive values were 82.9, 95.5, 98.9 and 52.5%, respectively. In specimens collected prospectively the RT-PCR sensitivity, specificity, positive and negative predictive values were 100, 87.9, 82.8 and 100%, respectively. There was complete concordance with subtype differentiation by hybridization probe melting temperature analysis and haemagglutination inhibition assay.

PMID:
15041206
DOI:
10.1016/j.jviromet.2003.12.005
[Indexed for MEDLINE]

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