Genotyping measles virus by real-time amplification refractory mutation system PCR represents a rapid approach for measles outbreak investigations.
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.
Waku-Kouomou
Diane
D
INSERM U404-Immunity and Vaccination, IFR128, 21 Avenue Tony Garnier, 69365 Lyon Cédex 03, France.
Alla
Amal
A
Blanquier
Bariza
B
Jeantet
Damien
D
Caidi
Hayat
H
Rguig
Ahmed
A
Freymuth
François
F
Wild
Fabian T
FT
D052160:Benzothiazoles; D003959:Diamines; D004196:Disease Outbreaks; D005456:Fluorescent Dyes; D005838:Genotype; D006801:Humans; D008457:Measles / Q000453:epidemiology* / Q000821:virology; D008459:Measles virus / Q000145:classification* / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D009930:Organic Chemicals; D016133:Polymerase Chain Reaction / Q000379:methods*; D011804:Quinolines; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA; D013997:Time Factors
D052160:Benzothiazoles; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I