Detection and identification of poliovirus in environmental samples using nucleic acid hybridization.
A procedure was developed to effectively extract viral RNA from poliovirus tissue-culture lysates while eliminating the hybridization background associated with tissue cultures uninfected with poliovirus. Poliovirus cDNA cloned into a pUC vector was used as probe. Both the recombinant plasmids and the cDNA showed great specificity towards poliovirus. However, both probes hybridized with the single-stranded DNA coliphage phi X174. Tissue culture was found to be an effective method to increase the number of viruses found in environmental samples to a level detectable by hybridization procedures, whereas direct hybridization of RNA from unamplified and highly concentrated raw wastewater showed poor hybridization signals. The specificity and sensitivity of the hybridization procedure developed during these studies indicate that this method may be best suited for the identification rather than the detection of viruses isolated from environmental samples.
Preston
D R
DR
Department of Microbiology and Cell Science, University of Florida, Gainesville 32611.
Chaudhry
G R
GR
Farrah
S R
SR
D002460:Cell Line; D015342:DNA Probes; D009500:Neutralization Tests; D009693:Nucleic Acid Hybridization; D017955:Poliovirus / Q000235:genetics / Q000302:isolation & purification*; D011237:Predictive Value of Tests; D012367:RNA, Viral / Q000032:analysis* / Q000302:isolation & purification; D014865:Waste Disposal, Fluid; D014871:Water Microbiology
D015342:DNA Probes; D012367:RNA, Viral