Rapid diagnosis of poliovirus infection by PCR amplification.
A single-tube, single-primer-set reverse transcription-PCR assay was developed for the rapid detection of polioviruses in infected tissue culture fluids and clinical materials. The poliovirus-specific PCR primers are located in the VP1-2A region of the poliovirus genome. They generate a 290-bp product and can be used in duplex reactions with general enterovirus primers. The primers span the region used for genotype determination, so that genotype analysis of wild-type polioviruses can be performed by direct sequencing of the PCR products. Of 125 virus isolates typed as polioviruses by neutralization assays, 125 (100%) were also positive by PCR, and of 38 isolates typed as non-polio enteroviruses by conventional techniques, 38 (100%) were also negative by PCR. The assay described here is rapid, highly sensitive, and specific and has clinical applicability in the diagnosis of poliovirus infections.
Chezzi
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Department of Virology, University of the Witwatersrand, Johannesburg, South Africa. claudiac@niv.ac.za
D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004770:Enterovirus / Q000235:genetics; D005243:Feces / Q000821:virology; D006801:Humans; D011051:Poliomyelitis / Q000175:diagnosis* / Q000821:virology*; D017955:Poliovirus / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013997:Time Factors
D017931:DNA Primers; D012367:RNA, Viral