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Comparative Study
. 2005 Jun;126(1-2):53-63.
doi: 10.1016/j.jviromet.2005.01.020.

Development of three multiplex RT-PCR assays for the detection of 12 respiratory RNA viruses

S Bellau-Pujol  1 , A Vabret, L Legrand, J Dina, S Gouarin, J Petitjean-Lecherbonnier, B Pozzetto, C Ginevra, F Freymuth
Affiliations
Comparative Study

Development of three multiplex RT-PCR assays for the detection of 12 respiratory RNA viruses

S Bellau-Pujol et al. J Virol Methods. 2005 Jun.

Abstract

Three multiplex hemi-nested RT-PCR assays were developed to detect simultaneously 12 RNA respiratory viruses: influenza viruses A, B and C, human respiratory syncytial virus (hRSV), human metapneumovirus (hMPV), parainfluenza virus types 1-4 (PIV-1, -2, -3 and -4), human coronavirus OC43 and 229E (HCoV) and rhinovirus (hRV). An internal amplification control was included in one of the RT-PCR assays. The RT-PCR multiplex 1 and the hemi-nested multiplex 1 detected 1 and 0.1 TCID50 of RSV A, respectively, and 0.01 and 0.001 TCID50 of influenza virus A/H3N2, respectively. Two hundred and three nasal aspirates from hospitalised children were retrospectively tested in comparison with two conventional methods: direct immunofluorescence assay and viral isolation technique. Almost all samples (89/91) that were positive by immunofluorescence assay and/or viral isolation technique were detected by the multiplex assay. This method also detected an additional 85 viruses and 33 co-infections. The overall sensitivity (98%), rapidity and enhanced efficiency of these multiplex hemi-nested RT-PCR assays suggest that they would be a significant improvement over conventional methods for the detection of a broad spectrum of respiratory viruses.

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Figures

Fig. 1
Fig. 1
Procedure and results of the multiplex RT-PCR and hemi-nested PCR.
See this image and copyright information in PMC

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