Real-time detection system for quantification of hepatitis C virus genome.
OBJECTIVE
For diagnosis of hepatitis C virus infection and monitoring of viral load in patients, a highly sensitive and accurate hepatitis C virus quantification system is essential.


METHODS
Hepatitis C virus genome was detected by real/time detection system using an ABI Prism 7700 sequence detector (Perkin Elmer Corp./Applied Biosystems, Foster City, CA).


RESULTS
As few as 10 copies of the genome were detected, and the quantification range was between 10(1) and 10(8) copies (r > 0.99). This system was 10-100-fold more sensitive than an Amplicor monitor (Roche Diagnostic Systems, Branchburg, NJ). The coefficient of variation values for both intra-assay precision and interassay reproducibility of identifying the genome quantification ranged from 0.37% to 2.00% and 0.88% to 4.66%, respectively. The system could detect the genome in 98% of patients with chronic hepatitis, 95.8% of patients with liver cirrhosis, and 100% of patients with hepatocellular carcinoma who had the antibody to hepatitis C virus, but could not detect the genome in patients without the antibody.


CONCLUSIONS
The establishment of a real-time detection system enables more accurate diagnosis of infection and monitoring of viral load in interferon-treated patients via quantification of viral genome.
Department of Microbiology, The Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan.
T
T
Takeuchi
A
A
Katsume
T
T
Tanaka
A
A
Abe
K
K
Inoue
K
K
Tsukiyama-Kohara
R
R
Kawaguchi
S
S
Tanaka
M
M
Kohara
D006528:Carcinoma, Hepatocellular / Q000821:virology; D016679:Genome, Viral; D005838:Genotype; D016174:Hepacivirus / Q000235:genetics* / Q000302:isolation & purification; D006526:Hepatitis C / Q000175:diagnosis*; D018937:Hepatitis C Antibodies / Q000097:blood; D006801:Humans; D008103:Liver Cirrhosis / Q000821:virology; D008113:Liver Neoplasms / Q000821:virology; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods*; D012367:RNA, Viral / Q000097:blood* / Q000235:genetics / Q000302:isolation & purification; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic
D018937:Hepatitis C Antibodies; D012367:RNA, Viral