Quantitative multiplex one-step RT-PCR assay for identification and quantitation of Sabin strains of poliovirus in clinical and environmental specimens.

ABSTRACT

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.