Mechnikov Research Institute for Vaccines and Sera, RAMS, 1st Dubrovskaya St. 15, Moscow, Russia.
J Virol Methods. 2013 Jan;187(1):57-64. doi: 10.1016/j.jviromet.2012.09.011. Epub 2012 Sep 24.
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.
在这项研究中，开发了一种基于 TaqMan 的实时逆转录聚合酶链反应（qPCR-RT）的快速定量方法，用于估计感染细胞培养上清液中麻疹、腮腺炎和风疹（MMR）病毒的滴度。qPCR-RT 测定法被证明是一种特异性、灵敏性、高效性和可重复性的方法。对于在不同感染复数下在 Vero 细胞中进行 MMR 病毒繁殖期间获得的 MMR 病毒样本，通过 qPCR-RT 测定法确定的滴度与通过传统的 MMR 病毒常用方法 - 50%细胞培养感染剂量（CCID(50)）测定法在配对样本中估计的感染性病毒进行了比较。在病毒接种后的特定时期内，Pearson 分析证明了两种方法之间存在显著相关性。此外，所建立的 qPCR-RT 测定法更快且不那么费力。该方法可用于替代或补充 MMR 疫苗生产过程中常规滴度估计的工具。
