Document Detail

Detection of hepatitis C virus genome in human serum by multi-targeted polymerase chain reaction.
MedLine Citation:
PMID:  8228939     Owner:  NLM     Status:  MEDLINE    
A multi-targeted "hemi-nested" PCR (M-PCR) assay in which the primer pairs derived from the 5' non-coding (5'NC) and the nonstructural protein 3 (NS3) regions of HCV genome were concurrently used for amplification in order to compare the sensitivity and specificity of polymerase chain reaction (PCR) with different primer pairs in detecting hepatitis C virus (HCV) genome. Sera from patients with virus-associated liver diseases were examined for the presence of HCV RNA by the M-PCR method following reverse transcription to cDNA. The amplified products derived from both the 5'NC and the NS3 regions were detected in 28 (70%) of the 40 HCV RNA-positive samples. However, 12 samples (30%) were devoid of the signal of NS3-derived product. Sensitivity tests using serial dilutions of HCV RNA revealed that the 5'NC-derived band was still detectable in the 10(5)-fold diluted sample by the M-PCR method, yet the NS3-derived band could hardly be detected in the 10(4)-fold diluted sample. Thus, as previously demonstrated by a single-targeted "nested" PCR assay, the present study using the M-PCR assay has clearly shown that the 5'NC-derived primers are more sensitive and specific than the NS3-derived primers in detecting HCV RNA.
M Okamoto; M Baba; E Kodama; K Sekine; T Takagi; R Kasukawa; S Shigeta
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of medical virology     Volume:  41     ISSN:  0146-6615     ISO Abbreviation:  J. Med. Virol.     Publication Date:  1993 Sep 
Date Detail:
Created Date:  1993-12-02     Completed Date:  1993-12-02     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7705876     Medline TA:  J Med Virol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  6-10     Citation Subset:  IM    
Department of Microbiology, Fukushima Medical College, Japan.
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MeSH Terms
BK Virus / classification*,  genetics,  isolation & purification
Base Sequence
Bone Marrow Transplantation
DNA, Viral / analysis,  urine
DNA-Directed DNA Polymerase / antagonists & inhibitors
Genome, Viral*
Kidney Transplantation
Molecular Sequence Data
Papillomavirus Infections / microbiology*
Polymerase Chain Reaction*
Sensitivity and Specificity
Taq Polymerase
Tumor Virus Infections / microbiology*
Urine / microbiology
Reg. No./Substance:
0/DNA, Viral; EC 2.7.7.-/Taq Polymerase; EC DNA Polymerase

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