| Title | Development of three multiplex RT-PCR assays for the detection of 12 respiratory RNA viruses. | ||
| Author | Bellau-Pujol, S; Vabret, A; Legrand, L; Dina, J; Gouarin, S; Petitjean-Lecherbonnier, J; Pozzetto, B; Ginevra, C; Freymuth, F | ||
| Journal | J Virol Methods | Publication Year/Month | 2005-Jun |
| PMID | 15847919 | PMCID | PMC7112904 |
| Affiliation | 1.Laboratory of Human and Molecular Virology, University Hospital, av. Georges Clemenceau, 14033 Caen, France. | ||
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.