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用于在临床标本中快速灵敏地鉴定和滴定脊髓灰质炎病毒的定量一步 RT-PCR 检测方法。

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

机构信息

Center for Biologics Evaluation and Research, US Food and Drug Administration, 1401 Rockville Pike, HFM 470, Rockville, MD 20852-1448, USA.

出版信息

J Virol Methods. 2013 Apr;189(1):7-14. doi: 10.1016/j.jviromet.2012.12.015. Epub 2013 Jan 7.

Abstract

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.

摘要

快速鉴定和定量检测临床标本中的脊髓灰质炎病毒对于监测和分析疫苗接种者的病毒脱落非常重要,这可用于评估黏膜免疫水平。本研究建立了一种用于鉴定和滴定三种脊髓灰质炎病毒血清型的一步定量 RT-PCR 方法。该方法可以替代基于细胞培养分离和随后确定脊髓灰质炎病毒血清型和病毒滴定的传统方法。该方法基于在同一管中进行定量 PCR 与逆转录反应。发现该多重检测方法可高度特异性、灵敏地定量检测三种血清型的脊髓灰质炎病毒,且仅需一天即可完成。

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