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Journal of Clinical Microbiology, February 2006, p. 487-494, Vol. 44, No. 2
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.2.487-494.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Genotyping Measles Virus by Real-Time Amplification Refractory Mutation System PCR Represents a Rapid Approach for Measles Outbreak Investigations

Diane Waku-Kouomou,^1,2,3 Amal Alla,⁴ Bariza Blanquier,^1,2 Damien Jeantet,^1,2 Hayat Caidi,⁴ Ahmed Rguig,⁵ François Freymuth,⁶ and Fabian T. Wild^1,2,3*

INSERM U404, F-69365, Lyon, France,¹ Université Claude Bernard Lyon, IFR128 BioScience Lyon-Gerland, F-69365 Lyon, France,² Centre National de Référence pour la Rougeole, Lyon, France,³ National Institut of Health, National Reference Laboratory for Measles, Rabat, Morocco,⁴ Direction d'épidémiologie et de lutte contre les maladies, service de la surveillance épidémiologique, Rabat, Morocco,⁵ Laboratoire de Virologie Humaine et Moléculaire, Hôpital Universitaire, Caen, France⁶

Received 1 June 2005/ Returned for modification 23 October 2005/ Accepted 3 November 2005

Real-time PCR has been developed to genotype measles virus (MV) isolates. MV strains circulating in epidemics in Gabon in 1984, Cameroon in 2001, Morocco in 2003, and France in 2004 were investigated. We developed a real-time amplification refractory mutation system PCR (RT-AMRS PCR) using SYBR green fluorescent dye. Six pairs of primers for RT-ARMS PCR were designed to specifically amplify genotypes A, B2, B3.1, B3.2, C2, and D7. Genotypes could be differentiated by melting curve analysis. All strains were also confirmed by direct sequencing. Using the result obtained by direct sequencing and phylogenetic analysis as the reference, the accuracy of MV by RT-ARMS PCR and melting curve analysis was 97%. However, the latter method is more rapid and sensitive than the former method. This method could be a useful tool for molecular epidemiological studies of MV, providing an efficient alternative for large-scale studies.

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* Corresponding author. Mailing address: INSERM U404—Immunity and Vaccination, IFR128, 21 Avenue Tony Garnier, 69365 Lyon Cédex 03, France. Phone: 33(0)4 37 28 23 92. Fax: 33(0)4 37 28 23 91. E-mail: wild{at}cervi-lyon.inserm.fr.

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Journal of Clinical Microbiology, February 2006, p. 487-494, Vol. 44, No. 2
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.2.487-494.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

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