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Journal of Clinical Microbiology, August 2003, p. 3631-3635, Vol. 41, No. 8
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.8.3631-3635.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Comparative Evaluation of Real-Time PCR Assays for Detection of the Human Metapneumovirus

Stéphanie Côté,1 Yacine Abed,1 and Guy Boivin1,2*

Infectious Diseases Research Center, Centre Hospitalier Universitaire de Québec (CHUQ-CHUL),1 Department of Medical Biology, Laval University, Québec City, Canada2

Received 16 January 2003/ Returned for modification 13 March 2003/ Accepted 5 May 2003

The human metapneumovirus (hMPV) is a new member of the Paramyxoviridae family associated with acute respiratory tract infections in humans. The objective of this study was to compare the sensitivity of real-time RT-PCR assays performed in a LightCycler instrument and designed to amplify the viral nucleoprotein (N), matrix (M), fusion (F), phosphoprotein (P), and polymerase (L) genes. In a first evaluation of 20 viral cultures with characteristics compatible with hMPV cytopathic effect, the PCR positivity rates were 100, 90, 75, 60, and 55% using primers for the N, L, M, P, and F genes. In a second evaluation of 10 nasopharyngeal aspirates from children with bronchiolitis and found to be positive for the hMPV N gene, the PCR positivity rates for the L, M, P, and F genes were 90, 60, 30, and 80%, respectively. The analytic sensitivity of the real-time RT-PCR assay for the hMPV N gene was 100 copies using a transcribed viral plasmid. In conclusion, real-time PCR assays aimed at amplifying the N and L genes which are coding for two internal viral proteins appear particularly suitable for hMPV diagnostic.

* Corresponding author. Mailing address: Centre de Recherche en Infectiologie, CHUQ, pavillon CHUL, room RC-709, 2705 blvd. Laurier, Ste-Foy, Québec, Canada G1V 4G2. Phone: (418) 654-2705. Fax: (418) 654-2715. E-mail: Guy.Boivin{at}crchul.ulaval.ca.

Journal of Clinical Microbiology, August 2003, p. 3631-3635, Vol. 41, No. 8
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.8.3631-3635.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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