Rapid identification of measles virus vaccine genotypes by real time PCR.

Roy F1, Mendoza L1, Hiebert J1, McNall RJ2, Bankamp B2, Connolly S2,3, Lüdde A4, Friedrich N4, Mankertz A4, Rota PA, Severini A5,6.

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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 RT-PCR method specific for genotype A measles virus (MeVA 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 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.