TY - JOUR
T1 - A novel method for liberating viral nucleic acid for assay of water samples with cDNA probes
AU - Richardson, Kenneth J.
AU - Margolin, Aaron B.
AU - Gerba, Charles P.
N1 - Funding Information:
This work was paid for in part by a grant from the United International Development.
PY - 1988/10
Y1 - 1988/10
N2 - 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°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.
AB - 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°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.
KW - Dot-blot hybridization
KW - Heat treatment
KW - Human placental RNasin
KW - Phenol-chloroform extraction
KW - Poliovirus
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U2 - 10.1016/0166-0934(88)90083-3
DO - 10.1016/0166-0934(88)90083-3
M3 - Article
C2 - 2848856
AN - SCOPUS:0024230469
VL - 22
SP - 13
EP - 21
JO - Journal of Virological Methods
JF - Journal of Virological Methods
SN - 0166-0934
IS - 1
ER -