title: Quantification of measles, mumps and rubella viruses using real-time quantitative TaqMan-based RT-PCR assay.
abstract: In this study, a rapid quantitative method using TaqMan-based real-time reverse transcription-polymerase chain reaction (qPCR-RT) has been developed for estimating the titers of measles, mumps and rubella (MMR) viruses in infected cell culture supernatants. The qPCR-RT assay was demonstrated to be a specific, sensitive, efficient and reproducible method. For MMR viral samples obtained during MMR viral propagations in Vero cells at a different multiplicity of infection, titers determined by the qPCR-RT assay have been compared with estimates of infectious virus obtained by a traditional commonly used method for MMR viruses - 50% cell culture infective dose (CCID(50)) assay, in paired samples. Pearson analysis evidenced a significant correlation between both methods for a certain period after viral inoculation. Furthermore, the established qPCR-RT assay was faster and less-laborious. The developed method could be used as an alternative method or a supplementary tool for the routine titer estimation during MMR vaccine production.
authors.affiliation: Mechnikov Research Institute for Vaccines and Sera, RAMS, 1st Dubrovskaya St. 15, Moscow, Russia. yulia.ammour@yahoo.fr
authors.forename: Y
authors.initials: Y
authors.lastname: Ammour
authors.forename: E
authors.initials: E
authors.lastname: Faizuloev
authors.forename: T
authors.initials: T
authors.lastname: Borisova
authors.forename: A
authors.initials: A
authors.lastname: Nikonova
authors.forename: G
authors.initials: G
authors.lastname: Dmitriev
authors.forename: S
authors.initials: S
authors.lastname: Lobodanov
authors.forename: V
authors.initials: V
authors.lastname: Zverev
mesh_terms: D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D006801:Humans; D008457:Measles / Q000175:diagnosis / Q000821:virology; D008459:Measles virus / Q000235:genetics / Q000302:isolation & purification*; D009107:Mumps / Q000175:diagnosis / Q000821:virology; D009109:Mumps virus / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012409:Rubella / Q000175:diagnosis / Q000821:virology; D012412:Rubella virus / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D014709:Vero Cells; D019562:Viral Load
chemical_list: D012367:RNA, Viral