Differentiation between vaccine-related and wild-type polioviruses using a heteroduplex mobility assay.
A heteroduplex mobility assay (HMA) was developed for intratypic differentiation between poliovirus isolates. The assay is based on polymerase chain reaction (PCR) amplification of a 480 base pair fragment which encodes a variable segment of VP1, followed by denaturation and reannealing of the resulting single strands with those from reference Sabin targets. Mismatches between wild-type and Sabin vaccine templates result in the formation of detectable heteroduplexes of reduced electrophoretic mobility. Poliovirus strains confirmed previously as wild-type or vaccine-like by PCR and sequencing were all correctly identified using the HMA. Mixtures of both wild-type and vaccine-like strains in a single isolate could also be detected using this technique. The results of this study demonstrate that heteroduplex analysis is a simple, rapid, and sensitive means for differentiating between vaccine-like and wild-type poliovirus isolates.
Chezzi
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National Institute for Virology, University of the Witwatersrand Medical School, Johannesburg, South Africa. claudiac@niv.ac.za
Schoub
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D002213:Capsid / Q000235:genetics; D036022:Capsid Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D009692:Nucleic Acid Heteroduplexes / Q000235:genetics*; D017955:Poliovirus / Q000235:genetics* / Q000302:isolation & purification*; D011055:Poliovirus Vaccine, Oral / Q000235:genetics* / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics* / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
D036022:Capsid Proteins; D009692:Nucleic Acid Heteroduplexes; D011055:Poliovirus Vaccine, Oral; D012367:RNA, Viral; C442540:VP1 protein, Poliovirus