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 leve l of healthy, HCV-negative individuals. Here, the sequence section from nt 57 to nt 328 within the 5'-NCR was assayed. Different combinations of prime rs were used in the nested PCR without a preceding reverse transcriptase st ep, resulting in fragments of the expected molecular weight size and also s horter and longer ones. It shows that the major part of the 5'-non-coding r egion (5'-NCR) of the hepatitis C virus genome is present in the DNA fracti on from peripheral blood mononuclear cells (PBMCs) of healthy, anti-HCV-neg ative individuals tested. Furthermore we designed experiments to prove the specificity of these findings, by using restriction enzymes for digest assa ys 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 par t of the internal ribosome entry site (IRES) structure of HCV at least is c ontained in the DNA of the individuals tested.