title: Highly sensitive detection of viral RNA genomes in blood specimens by an optimized reverse transcription-polymerase chain reaction.
abstract: A protocol was developed for a highly sensitive detection of viral RNA in blood specimens by reverse transcription coupled with a nested polymerase chain reaction. Using Japanese encephalitis virus (JEV) as a model, the optimized reverse transcription-polymerase chain reaction (ORTPCR) detects as few as 3-5 virions in 0.1 ml of whole blood specimens. The effectiveness of this assay system is confirmed by diagnosis of human hepatitis C viral (HCV) infection.
authors.affiliation: Kanonji Institute, Research Foundation for Microbial Diseases, Osaka University, Kagawa, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Murakami
authors.forename: Y
authors.initials: Y
authors.lastname: Takahashi
authors.forename: S
authors.initials: S
authors.lastname: Yoshida
authors.forename: I
authors.initials: I
authors.lastname: Fuke
authors.forename: K
authors.initials: K
authors.lastname: Ohmae
authors.forename: C
authors.initials: C
authors.lastname: Mori
authors.forename: M
authors.initials: M
authors.lastname: Takagi
authors.forename: A
authors.initials: A
authors.lastname: Takamizawa
authors.forename: H
authors.initials: H
authors.lastname: Okayama
mesh_terms: D000818:Animals; D001483:Base Sequence; D002648:Child; D018349:Encephalitis Viruses, Japanese / Q000302:isolation & purification; D016174:Hepacivirus / Q000302:isolation & purification; D006526:Hepatitis C / Q000382:microbiology; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000097:blood*; D012194:RNA-Directed DNA Polymerase; D012680:Sensitivity and Specificity
chemical_list: D012367:RNA, Viral; D012194:RNA-Directed DNA Polymerase