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ID 32281
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Author
Nozaki, Akito
Naganuma, Atsushi
Nakamura, Takashi
Tanaka, Katsuaki
Sekihara, Hisahiko
Abstract

We have developed a reliable internally controlled RT-nested PCR method for the detection of hepatitis C virus (HCV) RNA using in vitro synthesized Renilla luciferase (Rluc) RNA as an internal control. Using this method, the 5'-noncoding region of HCV RNA (144 nucleotides) and Rluc RNA (276 nucleotides) were efficiently amplified in a single tube, and the sensitivity and specificity of this method were comparable to standard RT-nested PCR. This method was successfully performed on RNA specimens obtained from in vitro HCV-infected human hepatocyte PH5CH8 cells, which support HCV replication. In addition, we demonstrated that this method was useful for the evaluation of antiviral reagents by confirming the anti-HCV activity of bovine lactoferrin, which we previously found to be a new inhibitor of HCV infection. Therefore, this method may be useful for the studies of not only HCV but also of other viruses.

Keywords
Hepatitis C virus
Reverse transcriptionnested PCR (RT-nested PCR)
internal control
Amo Type
Article
Published Date
2000-12
Publication Title
Acta Medica Okayama
Volume
volume54
Issue
issue6
Publisher
Okayama University Medical School
Start Page
253
End Page
257
ISSN
0386-300X
NCID
AA00508441
Content Type
Journal Article
language
英語
File Version
publisher
Refereed
True
PubMed ID
Web of Science KeyUT