Isolation of hepatitis C virus RNA from serum for reverse transcription-PCR.
Standard multistep extraction and isolation of RNA for hepatitis C virus (HCV) reverse transcription (RT)-PCR are impractical for routine use in clinical laboratories. We compared three simple commercially available methods for RNA isolation (RNAzol B, TRISOLV, and ULTRASPEC; Biotecx Laboratories, Houston, Tex.) and a total nucleic acid isolation method (IsoQuick; MicroProbe Corp., Garden Grove, Calif.) for the recovery of HCV RNA from sera obtained from 12 viremic patients for RT-PCR. RNAzol B, TRISOLV, ULTRASPEC, and IsoQuick extraction methods detected 87.5, 79.2, 33.3, and 58.3% of the paired positive samples, respectively. The method used for isolation of RNA is an important concern when optimizing HCV RT-PCR.
Nolte
F S
FS
Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, Georgia 30322.
Thurmond
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C
Mitchell
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PS
D005069:Evaluation Studies as Topic; D016174:Hepacivirus / Q000235:genetics* / Q000302:isolation & purification*; D006526:Hepatitis C / Q000175:diagnosis / Q000382:microbiology; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012367:RNA, Viral / Q000097:blood* / Q000235:genetics*; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic; D014766:Viremia / Q000175:diagnosis / Q000382:microbiology
D012367:RNA, Viral