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We developed a novel single-step reverse transcription-polymerase chain reaction (RT-PCR), which is equal in sensitivity and specificity to RT-nested PCR, based on both reverse transcriptase and Taq DNA polymerase working efficiently under single buffer reaction conditions. Using in vitro synthesized hepatitis C virus (HCV) RNA, it was demonstrated that 10-100 copies of HCV RNA could be detected with a set of primers that amplify a 144 base-pair sequence unique to the 5'-noncoding region of HCV RNA. Furthermore, this method was successfully performed on serum and liver biopsy specimens obtained from patients with chronic hepatitis C. In addition, HCV RNA from in vitro HCV-infected MT-2C cells, which supported HCV replication, was also detected by this method. The method is anticipated to improve the detection of small amounts of RNA, such as that of HCV, promoting both labor savings and the prevention of carry-over contamination.
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
June 1995,
Journal of virological methods,
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
August 1993,
Hepatology (Baltimore, Md.),
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
May 1997,
Journal of medical virology,
T Mizutani, and
M Ikeda, and
S Saito, and
K Sugiyama, and
K Shimotohno, and
N Kato
June 1997,
Journal of virological methods,
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,
