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Journal of Clinical Microbiology, February 2005, p. 589-595, Vol. 43, No. 2
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.2.589-595.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Evaluation of a Multiplex Real-Time Reverse Transcriptase PCR Assay for Detection and Differentiation of Influenza Viruses A and B during the 2001-2002 Influenza Season in Israel

Musa Hindiyeh,1 Virginia Levy,1 Roberto Azar,1 Noemi Varsano,1 Liora Regev,1 Yael Shalev,1 Zehava Grossman,1 and Ella Mendelson1,2*

Central Virology Laboratory, Public Health Services, Ministry of Health, Chaim Sheba Medical Center, Tel-Hashomer,1 Faculty of Life Sciences, Bar Ilan University, Ramat-Gan, Israel2

Received 12 August 2004/ Returned for modification 26 August 2004/ Accepted 14 October 2004

The ability to rapidly diagnose influenza virus infections is of the utmost importance in the evaluation of patients with upper respiratory tract infections. It is also important for the influenza surveillance activities performed by national influenza centers. In the present study we modified a multiplex real-time reverse transcriptase PCR (RT-PCR) assay (which uses TaqMan chemistry) and evaluated it for its ability to detect and concomitantly differentiate influenza viruses A and B in 370 patient samples collected during the 2001-2002 influenza season in Israel. The performance of the TaqMan assay was compared to those of a multiplex one-step RT-PCR with gel detection, a shell vial immunofluorescence assay, and virus isolation in tissue culture. The TaqMan assay had an excellent sensitivity for the detection of influenza viruses compared to that of tissue culture. The overall sensitivity and specificity of the TaqMan assay compared to the results of culture were 98.4 and 85.5%, respectively. The sensitivity and specificity of the TaqMan assay for the detection of influenza virus A alone were 100 and 91.1%, respectively. On the other hand, the sensitivity and specificity for the detection of influenza virus B alone were 95.7 and 98.7%, respectively. The rapid turnaround time for the performance of the TaqMan assay (4.5 h) and the relatively low direct cost encourage the routine use of this assay in place of tissue culture. We conclude that the multiplex TaqMan assay is highly suitable for the rapid diagnosis of influenza virus infections both in well-established molecular biology laboratories and in reference clinical laboratories.

* Corresponding author. Mailing address: Central Virology Laboratory, Public Health Services, Ministry of Health, Chaim Sheba Medical Center, Tel-Hashomer 52621, Israel. Phone: 3-530-2421. Fax: 3-530-2457. E-mail: ellamen{at}sheba.health.gov.il.

Journal of Clinical Microbiology, February 2005, p. 589-595, Vol. 43, No. 2
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.2.589-595.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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