A comparison between the phenol-chloroform method of RNA extraction and the QIAamp viral RNA kit in the extraction of hepatitis C and GB virus-C/hepatitis G viral RNA from serum.
Viral RNAs are detected commonly in serum by hybridization and polymerase chain reaction (PCR) methods for both clinical and research purposes. Two methods of extracting viral RNA were evaluated prior to amplification by PCR. One was a conventional phenol-chloroform method and the other used a standardized, manufactured kit. The efficiency of extraction was tested by semi-quantitative amplification of hepatitis C viral and GB virus-C/hepatitis G viral RNAs. The standarized commercial method, although more time efficient, resulted in an approximately ten fold less sensitivity. Thus, in situations where maximum sensitivity is needed, the more labor intenstive phenol choloroform method is recommended.
Fanson
B G
BG
Department of Internal Medicine, Saint Louis University School of Medicine, 1402 South Grand Bld. 63104, St Louis, MO, USA.
Osmack
P
P
Di Bisceglie
A M
AM
D002725:Chloroform; D018067:Flaviviridae / Q000302:isolation & purification*; D016174:Hepacivirus / Q000302:isolation & purification*; D006801:Humans; D019800:Phenol; D012367:RNA, Viral / Q000032:analysis / Q000097:blood / Q000302:isolation & purification*; D011933:Reagent Kits, Diagnostic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic; D019800:Phenol; D002725:Chloroform