Hepatitis C virus-specific DNA sequences in human DNA: differentiation by means of restriction enzyme analysis at the DNA level in healthy, anti-HCV-negative individuals.
This study aimed to look for further HCV-specific sequences at the DNA level of healthy, HCV-negative individuals. Here, the sequence section from nt 57 to nt 328 within the 5'-NCR was assayed. Different combinations of primers were used in the nested PCR without a preceding reverse transcriptase step, resulting in fragments of the expected molecular weight size and also shorter and longer ones. It shows that the major part of the 5'-non-coding region (5'-NCR) of the hepatitis C virus genome is present in the DNA fraction from peripheral blood mononuclear cells (PBMCs) of healthy, anti-HCV-negative individuals tested. Furthermore we designed experiments to prove the specificity of these findings, by using restriction enzymes for digest assays of the target DNA before PCR (pre-PCR digest) and of the products after PCR (post-PCR digest). In conclusion, our study indicates that the main part of the internal ribosome entry site (IRES) structure of HCV at least is contained in the DNA of the individuals tested.
Dennin
R H
RH
Institut für Medizinische Mikrobiologie und Hygiene, Medizinische Universität zu Lübeck, Germany. dennin@hygiene.MU-luebeck.de
Wo
J
J
Chen
Z
Z
D001483:Base Sequence; D002478:Cells, Cultured; D004279:DNA, Viral / Q000032:analysis* / Q000097:blood; D005260:Female; D006367:HeLa Cells; D016174:Hepacivirus / Q000235:genetics*; D006526:Hepatitis C / Q000097:blood / Q000276:immunology; D018937:Hepatitis C Antibodies / Q000097:blood*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping / Q000379:methods; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
D004279:DNA, Viral; D018937:Hepatitis C Antibodies