Measles and rubella virus cause fever/rash diseases that are difficult to differentiate clinically. Both viruses can be detected in the same clinical specimens and are propagated on the same cell cultures. A single-tube multiplex TaqMan assay is described for the simultaneous and rapid detection of the full spectrum of known genetic variants. The performance of the assay is similar to a conventional nested PCR and generates cDNA with random primers which can be used directly for virus genotyping.