title: Rapid Identification of Measles Virus Vaccine Genotype by Real-Time PCR.
abstract: During measles outbreaks, it is important to be able to rapidly distinguish between measles cases and vaccine reactions to avoid unnecessary outbreak response measures such as case isolation and contact investigations. We have developed a real-time reverse transcription-PCR (RT-PCR) method specific for genotype A measles virus (MeV) (MeVA RT-quantitative PCR [RT-qPCR]) that can identify measles vaccine strains rapidly, with high throughput, and without the need for sequencing to determine the genotype. We have evaluated the method independently in three measles reference laboratories using two platforms, the Roche LightCycler 480 system and the Applied Biosystems (ABI) 7500 real-time PCR system. In comparison to the standard real-time RT-PCR method, the MeVA RT-qPCR showed 99.5% specificity for genotype A and 94% sensitivity for both platforms. The new assay was able to detect RNA from five currently used vaccine strains, AIK-C, CAM-70, Edmonston-Zagreb, Moraten, and Shanghai-191. The MeVA RT-qPCR assay has been used successfully for measles surveillance in reference laboratories, and it could be readily deployed to national and subnational laboratories on a wide scale.
authors.affiliation: Viral Exanthemata and STD Section, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.
authors.forename: Felicia
authors.initials: F
authors.lastname: Roy
authors.affiliation: Viral Exanthemata and STD Section, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.
authors.forename: Lillian
authors.initials: L
authors.lastname: Mendoza
authors.affiliation: Viral Exanthemata and STD Section, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.
authors.forename: Joanne
authors.initials: J
authors.lastname: Hiebert
authors.affiliation: Division of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
authors.forename: Rebecca J
authors.initials: RJ
authors.lastname: McNall
authors.affiliation: Division of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
authors.forename: Bettina
authors.initials: B
authors.lastname: Bankamp
authors.affiliation: Division of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.|Emory University, Atlanta, Georgia, USA.
authors.forename: Sarah
authors.initials: S
authors.lastname: Connolly
authors.affiliation: Unit Measles, Mumps, Rubella, and Viruses Affecting Immunocompromised Patients, Robert Koch Institute, Berlin, Germany.
authors.forename: Amy
authors.initials: A
authors.lastname: Lüdde
authors.affiliation: Unit Measles, Mumps, Rubella, and Viruses Affecting Immunocompromised Patients, Robert Koch Institute, Berlin, Germany.
authors.forename: Nicole
authors.initials: N
authors.lastname: Friedrich
authors.affiliation: Unit Measles, Mumps, Rubella, and Viruses Affecting Immunocompromised Patients, Robert Koch Institute, Berlin, Germany.
authors.forename: Annette
authors.initials: A
authors.lastname: Mankertz
authors.affiliation: Division of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Rota
authors.affiliation: Viral Exanthemata and STD Section, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada alberto.severini@phac-aspc.gc.ca.|Dept. of Medical Microbiology, Faculty of Health Sciences, University of Manitoba, Winnipeg, Manitoba, Canada.
authors.forename: Alberto
authors.initials: A
authors.lastname: Severini
mesh_terms: D005838:Genotype; D006801:Humans; D008458:Measles Vaccine / Q000235:genetics*; D008459:Measles virus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D008458:Measles Vaccine
keywords: PCR; genotyping; measles; measles vaccine; molecular methods