用于鉴定和定量临床和环境标本中脊髓灰质炎病毒 Sabin 株的定量多重一步 RT-PCR 检测方法。
Quantitative multiplex one-step RT-PCR assay for identification and quantitation of Sabin strains of poliovirus in clinical and environmental specimens.
机构信息
Center for Biologics Evaluation and Research, US Food and Drug Administration, 10903 New Hampshire Avenue, Silver Spring, MD 20993, United States.
Fighting Infectious Disease in Emerging Countries (FIDEC) Miami, Fl 33145, United States.
出版信息
J Virol Methods. 2018 Sep;259:74-80. doi: 10.1016/j.jviromet.2018.06.009. Epub 2018 Jun 18.
An improved quantitative multiplex one-step RT-PCR (qmosRT-PCR) for simultaneous identification and quantitation of all three serotypes of poliovirus is described. It is based on using serotype-specific primers and fluorescent TaqMan oligonucleotide probes. The assay can be used for high-throughput screening of samples for the presence of poliovirus, poliovirus surveillance and for evaluation of virus shedding by vaccine recipients in clinical trials to assess mucosal immunity. It could replace conventional methods based on cell culture virus isolation followed by serotyping. The assay takes only few hours, and was found to be simple, specific, sensitive and has large quantitative linearity range. In addition, the method could be used as readout in PCR-based poliovirus titration and neutralization assays.
一种改良的定量多重一步 RT-PCR（qmosRT-PCR）方法，可用于同时鉴定和定量所有三种血清型脊髓灰质炎病毒。它基于使用血清型特异性引物和荧光 TaqMan 寡核苷酸探针。该检测方法可用于高通量筛选样本中脊髓灰质炎病毒的存在，进行脊髓灰质炎病毒监测，并评估临床试验中疫苗接种者的病毒脱落情况，以评估黏膜免疫。它可以替代基于细胞培养病毒分离和随后的血清分型的传统方法。该检测方法仅需几个小时，并且被发现简单、特异、敏感，具有较大的定量线性范围。此外，该方法可用于基于 PCR 的脊髓灰质炎病毒滴定和中和测定中。
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