title: Detection of hepatitis C virus RNA in serum, by combining reverse transcription and polymerase chain reaction in one tube.
abstract: 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.
authors.affiliation: North Manchester Virus Laboratory, Booth Hall Children's Hospital, Manchester, UK.
authors.forename: P
authors.initials: P
authors.lastname: Tilston
authors.forename: G
authors.initials: G
authors.lastname: Corbitt
mesh_terms: 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
chemical_list: D018076:DNA, Complementary; D019384:Nucleic Acid Synthesis Inhibitors; D012367:RNA, Viral; D014885:Waxes; D019914:Taq Polymerase; D012194:RNA-Directed DNA Polymerase