Highly sensitive detection of viral RNA genomes in blood specimens by an optimized reverse transcription-polymerase chain reaction.
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.
Murakami
S
S
Kanonji Institute, Research Foundation for Microbial Diseases, Osaka University, Kagawa, Japan.
Takahashi
Y
Y
Yoshida
S
S
Fuke
I
I
Ohmae
K
K
Mori
C
C
Takagi
M
M
Takamizawa
A
A
Okayama
H
H
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
D012367:RNA, Viral; D012194:RNA-Directed DNA Polymerase