Quantitative one-step RT-PCR assay for rapid and sensitive identification and titration of polioviruses in clinical specimens

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• 作者：Majid Laassri ; ^{majid.laassri@fda.hhs.gov} ; Anthony DiPiazza ; Bella Bidzhieva ; Tatiana Zagorodnyaya ; Konstantin Chumakov
• 关键词：Real-time PCR ; Quantitative PCR ; Poliovirus surveillance ; Poliovirus excretion ; Shedding ; Mucosal immunity ; Titration
• 刊名：Journal of Virological Methods
• 出版年：2013
• 出版时间：April, 2013
• 年：2013
• 卷：189
• 期：1
• 页码：7-14
• 全文大小：560 K

文摘

Rapid identification and quantitation of polioviruses in clinical specimens is important for surveillance and analysis of virus shedding by vaccine recipients, which could be used to assess the level of mucosal immunity. A quantitative one step RT-PCR was developed for identification and titration of all three poliovirus serotypes. The assay could be an alternative to the traditional procedure based on cell culture isolation and subsequent determination of poliovirus serotype and virus titration. The method is based on quantitative PCR performed with reverse transcription reaction in the same tube. The multiplex assay that quantifies all three serotypes of poliovirus was found to be highly specific, sensitive, and takes only one day to complete.