title: Potency estimation of measles, mumps and rubella trivalent vaccines with quantitative PCR infectivity assay.
abstract: The quantitative PCR infectivity assay is a combination of virus propagation and quantitative PCR. Previously [Schalk JAC, van den Elzen C, Ovelgonne H, Baas C, Jongen PMJM. Estimation of the number of infectious measles viruses in live virus vaccines using quantitative real-time PCR. J Virol Methods 2004;117:179-87.], we used this assay to estimate the titer of infectious measles virus in trivalent, live, measles, mumps, rubella vaccines (MMR). Here we describe the further improvement and development of the assay for simultaneous potency estimation of measles, mumps and rubella viruses. The potency of measles and mumps virus is estimated within one assay after 1 day of cell culture. The potency of rubella virus is estimated in a separate assay after 2 days of cell culture. Compared to conventional CCID50 and plaque assays, the quantitative PCR infectivity assay has the advantage in being fast because the assay is not dependent on the formation of cytopathic effect. Furthermore assay design is simplified: serological neutralization can be omitted because PCR is virus-specific and, under the conditions used, the individual components of trivalent measles, mumps, rubella vaccines do not interfere with each other. The assay was validated and compared to the performance of a plaque assay.
authors.affiliation: National Institute for Public Health and the Environment, Centre for Biological Medicines and Medical Technology, Antonie van Leeuwenhoeklaan 9, 3721 MA Bilthoven, The Netherlands. marjolijn.schalk@rivm.nl
authors.forename: J A C
authors.initials: JA
authors.lastname: Schalk
authors.forename: C G J C A
authors.initials: CG
authors.lastname: de Vries
authors.forename: P M J M
authors.initials: PM
authors.lastname: Jongen
mesh_terms: D000818:Animals; D001483:Base Sequence; D002522:Chlorocebus aethiops; D017931:DNA Primers; D008459:Measles virus / Q000472:pathogenicity / Q000502:physiology; D022542:Measles-Mumps-Rubella Vaccine / Q000235:genetics / Q000276:immunology*; D009109:Mumps virus / Q000472:pathogenicity / Q000502:physiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics; D015203:Reproducibility of Results; D012412:Rubella virus / Q000472:pathogenicity / Q000502:physiology; D014709:Vero Cells; D014774:Virulence; D014779:Virus Replication
chemical_list: D017931:DNA Primers; D022542:Measles-Mumps-Rubella Vaccine; D012367:RNA, Viral