A novel method for liberating viral nucleic acid for assay of water samples with cDNA probes.
Rapid and sensitive methods are needed for the detection of enteric viruses to ensure proper drinking water quality. Gene probes have been shown to be useful for this purpose. Previously, samples to be assayed were treated with a series of phenol-chloroform extractions to release the viral nucleic acid. We have developed a more rapid procedure for liberating or exposing the genome of poliovirus for probing. In this study, a poliovirus model was used to test the ability of heat (65 degrees C for 30 min) for release or exposure of viral nucleic acid. Several different RNase inhibitors were tested for their ability to prevent viral RNA degradation. A comparison of the two methods indicates phenol-chloroform extraction is not necessary before probing. In addition to saving 2-4 h of time, maximum sensitivity levels were consistently obtained using this novel procedure.
Richardson
K J
KJ
Department of Microbiology and Immunology, University of Arizona, Tucson 85721.
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D004247:DNA; D015342:DNA Probes; D004770:Enterovirus / Q000302:isolation & purification; D006358:Hot Temperature; D012367:RNA, Viral / Q000302:isolation & purification*; D012260:Ribonucleases / Q000037:antagonists & inhibitors; D014871:Water Microbiology
D015342:DNA Probes; D012367:RNA, Viral; D004247:DNA; D012260:Ribonucleases