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We developed a novel single-tube reverse transcription-polymerase chain reaction (RT-PCR) for the specific detection of negative-stranded hepatitis C virus (HCV) RNA. By using in vitro synthesized positive- and negative-stranded HCV RNAs, it was demonstrated that as few as 50 copies of negative-stranded HCV RNA could be specifically detected with a set of primers that amplify a 232-base pair sequence unique to the 5'-non-coding region of HCV RNA, while 10(8) copies of positive-stranded HCV RNA were not detected. In addition, we demonstrated that this method allows the detection as few as 100 copies of negative-stranded HCV RNA even with the coexistence of a 100-fold excess of positive-stranded HCV RNA. Furthermore, with this method, negative-stranded HCV RNA was detected in RNAs from liver biopsy specimens obtained from patients with chronic hepatitis C, but not in RNAs from HCV-positive sera.
T Mizutani, and
M Ikeda, and
S Saito, and
K Sugiyama, and
K Shimotohno, and
N Kato
August 2000,
Hepatology (Baltimore, Md.),
T Mizutani, and
M Ikeda, and
S Saito, and
K Sugiyama, and
K Shimotohno, and
N Kato
September 1995,
Nihon rinsho. Japanese journal of clinical medicine,
T Mizutani, and
M Ikeda, and
S Saito, and
K Sugiyama, and
K Shimotohno, and
N Kato
January 1999,
Clinical chemistry and laboratory medicine,
T Mizutani, and
M Ikeda, and
S Saito, and
K Sugiyama, and
K Shimotohno, and
N Kato
November 1992,
European journal of clinical chemistry and clinical biochemistry : journal of the Forum of European Clinical Chemistry Societies,
T Mizutani, and
M Ikeda, and
S Saito, and
K Sugiyama, and
K Shimotohno, and
N Kato
April 1993,
Journal of clinical microbiology,
T Mizutani, and
M Ikeda, and
S Saito, and
K Sugiyama, and
K Shimotohno, and
N Kato
January 1995,
Analytical biochemistry,
T Mizutani, and
M Ikeda, and
S Saito, and
K Sugiyama, and
K Shimotohno, and
N Kato
May 1997,
Journal of medical virology,
T Mizutani, and
M Ikeda, and
S Saito, and
K Sugiyama, and
K Shimotohno, and
N Kato
January 1995,
Research in virology,
T Mizutani, and
M Ikeda, and
S Saito, and
K Sugiyama, and
K Shimotohno, and
N Kato
March 1992,
Hepatology (Baltimore, Md.),
T Mizutani, and
M Ikeda, and
S Saito, and
K Sugiyama, and
K Shimotohno, and
N Kato
September 1998,
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology,
