Detection of hepatitis C virus RNA in serum, by combining reverse transcription and polymerase chain reaction in one tube.
Reverse transcription of viral RNA into cDNA and its subsequent amplification by polymerase chain reaction (PCR) are generally carried out as two separate reactions. Here we describe a novel method for the detection of HCV RNA in serum combining both steps in one reaction tube using a wax interface to separate the two sets of reactants during initial cDNA synthesis. This enabled optimisation of both reactions, simplified the test and thereby reduced the risk of cross-contamination.
Tilston
P
P
North Manchester Virus Laboratory, Booth Hall Children's Hospital, Manchester, UK.
Corbitt
G
G
D055598:Chemical Phenomena; D002627:Chemistry, Physical; D018076:DNA, Complementary / Q000032:analysis / Q000235:genetics; D016174:Hepacivirus / Q000302:isolation & purification*; D006526:Hepatitis C / Q000382:microbiology*; D019384:Nucleic Acid Synthesis Inhibitors; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D012367:RNA, Viral / Q000097:blood* / Q000302:isolation & purification; D012194:RNA-Directed DNA Polymerase; D012680:Sensitivity and Specificity; D019914:Taq Polymerase; D013696:Temperature; D014766:Viremia / Q000382:microbiology*; D014885:Waxes
D018076:DNA, Complementary; D019384:Nucleic Acid Synthesis Inhibitors; D012367:RNA, Viral; D014885:Waxes; D019914:Taq Polymerase; D012194:RNA-Directed DNA Polymerase