Detection and rapid differentiation of human enteroviruses in water sources by restriction enzyme analysis.
The objective of this study was to assess the application and efficiency of molecular techniques for the detection and serotyping of enteroviruses from environmental water samples. Samples of water were collected at regular intervals upstream and downstream of an informal settlement. Techniques for the detection of enteroviruses included a reverse transcription polymerase chain reaction (RT-PCR), nested PCR (n-PCR) and Sabin-specific triplex PCR. A specific 297 bp fragment was amplified by the n-PCR and subjected to restriction enzyme (RE) analysis to differentiate between various serotypes of prototypical enteroviruses. Enteroviruses that gave inconclusive restriction patterns were typed by partial sequencing of the VP1 region. Results indicated a high incidence of enteroviruses, predominantly coxsackie B viruses. The results on polioviruses, as well as other enteroviruses, contributed valuable information on enteroviruses circulating in the community. The molecular approach described here proved suitable for the rapid, sensitive, specific and cost effective, simultaneous detection and typing of enteroviruses in water.
Vivier
J C
JC
Dept of Medical Virology, University of Pretoria, P.O. Box 2034, Pretoria 0001, South Africa. jvivier@medic.up.ac.za
Clay
C G
CG
Grabow
W O
WO
D003365:Costs and Cost Analysis; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D004770:Enterovirus / Q000235:genetics*; D004784:Environmental Monitoring / Q000379:methods; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D012703:Serotyping; D014876:Water Pollution / Q000032:analysis*
D004279:DNA, Viral