Detection and quantitation of hepatitis C virus RNA in serum using the polymerase chain reaction and a colorimetric enzymatic detection system.
A sensitive, non-isotopic method for detecting and quantifying hepatitis C virus (HCV) RNA in serum using the reverse transcriptase-polymerase chain reaction (RT-PCR) and a hybridization specific, colorimetric biotin-avidin peroxidase detection system has been developed. The sensitivity of the PCR-colorimetric system was determined using RNA synthesized from cloned HCV cDNA. The assay could detect as few as 10 molecules of HCV RNA, comparable to the sensitivity achieved with double PCR using nested primers. Thus, this colorimetric assay can detect low levels of HCV RNA in serum and appears to be quantitative, suggesting that this technique may be applied to rapid screening of large numbers of samples and to monitor the effect of antiviral therapy.
Shindo
M
M
Hepatitis Studies Section, NIH, NIDDK, Bethesda, MD.
Di Bisceglie
A M
AM
Silver
J
J
Limjoco
T
T
Hoofnagle
J H
JH
Feinstone
S M
SM
D000818:Animals; D001360:Avidin; D001483:Base Sequence; D001710:Biotin; D003124:Colorimetry; D018076:DNA, Complementary / Q000032:analysis; D004587:Electrophoresis, Agar Gel; D016174:Hepacivirus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D006526:Hepatitis C / Q000821:virology*; D008969:Molecular Sequence Data; D002679:Pan troglodytes / Q000097:blood / Q000821:virology; D009195:Peroxidase; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000097:blood*; D012680:Sensitivity and Specificity; D014766:Viremia / Q000821:virology*
D018076:DNA, Complementary; D012367:RNA, Viral; D001360:Avidin; D001710:Biotin; D009195:Peroxidase