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Comparative Study
. 2009 Mar;47(3):527-32.
doi: 10.1128/JCM.01213-08. Epub 2009 Jan 7.

Comparison of viral isolation and multiplex real-time reverse transcription-PCR for confirmation of respiratory syncytial virus and influenza virus detection by antigen immunoassays

R S Liao  1 , L L Tomalty, A Majury, D E Zoutman
Affiliations
Comparative Study

Comparison of viral isolation and multiplex real-time reverse transcription-PCR for confirmation of respiratory syncytial virus and influenza virus detection by antigen immunoassays

R S Liao et al. J Clin Microbiol. 2009 Mar.

Abstract

We evaluated the Prodesse ProFlu-1 real-time reverse transcription-PCR multiplex assay with the SmartCycler instrument for the detection of human respiratory syncytial virus (RSV) and influenza A and B viruses in comparison to conventional cell culture and antigen immunoassays with the BD Directigen A+B and Binax NOW RSV assays over two successive respiratory virus seasons. Ninety-two percent of the 361 specimens tested were nasopharyngeal aspirates obtained from individual patients, of which 119 were positive for RSV and 59 were positive for influenza virus. The median age of the patients whose specimens were positive for RSV and influenza virus were 6.3 months and 42.4 years, respectively. The specificity of all of the methods tested was >or=99%, and the individual sensitivities of NOW RSV, RSV culture, Directigen A+B, influenza virus culture, and the Proflu-1 PCR for influenza/RSV were 82% (95% confidence interval [CI], 73 to 88), 57% (95% CI, 44 to 69), 59% (95% CI, 44 to 72), 54% (95% CI, 38 to 69), and 98% (95% CI, 93 to 100)/95% (95% CI, 85 to 99), respectively. In a clinical setting where viral isolation is performed to confirm rapid antigen immunoassay results for these common respiratory viruses, one-step real-time reverse transcriptase PCR testing can be a more sensitive and timely confirmatory method.

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Figures

FIG. 1.
FIG. 1.
Comparison of PCR CT values obtained with the Proflu-1 multiplex assay for specimens that tested positive or negative for RSV (A) or influenza virus (B) by virus isolation or antigen immunoassay. The differences between the mean PCR CT values of antigen (Ag)-positive and -negative specimens were significant (NOW RSV, P < 0.0001; Directigen A+B, P = 0.0235).
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