Document Detail


A reliable internally controlled RT-nested PCR method for the detection of hepatitis C virus RNA.
MedLine Citation:
PMID:  11132918     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
A Nozaki; A Naganuma; T Nakamura; K Tanaka; H Sekihara; N Kato
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Acta medica Okayama     Volume:  54     ISSN:  0386-300X     ISO Abbreviation:  Acta Med. Okayama     Publication Date:  2000 Dec 
Date Detail:
Created Date:  2000-12-29     Completed Date:  2001-03-08     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0417611     Medline TA:  Acta Med Okayama     Country:  Japan    
Other Details:
Languages:  eng     Pagination:  253-7     Citation Subset:  IM    
Affiliation:
Department of Molecular Biology, Institute of Cellular and Molecular Biology, Okayama University Medical School, Japan.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antiviral Agents / pharmacology
Beetles / enzymology
Cattle
Cell Line
Hepacivirus / drug effects,  genetics*
Hepatocytes / chemistry,  virology
Humans
Lactoferrin / pharmacology
Luciferases / genetics
RNA
RNA, Viral / analysis*
Reverse Transcriptase Polymerase Chain Reaction*
Sensitivity and Specificity
Virus Replication
Chemical
Reg. No./Substance:
0/Antiviral Agents; 0/Lactoferrin; 0/RNA, Viral; 63231-63-0/RNA; EC 1.13.12.-/Luciferases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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