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首页> 外文期刊>Clinical chemistry and laboratory medicine: CCLM >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.
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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.

机译：人类DNA中的丙型肝炎病毒特异性DNA序列：在健康的抗HCV阴性个体中通过限制酶分析在DNA水平上进行分化。

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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.
机译：这项研究的目的是在健康的HCV阴性个体的DNA水平上寻找进一步的HCV特异性序列。在此，分析了5'-NCR内从nt 57至nt 328的序列部分。巢式PCR中使用了不同的引物组合，而没有前面的逆转录酶步骤，从而产生了具有预期分子量大小的片段，并且片段的长度越来越短。结果表明，丙型肝炎病毒基因组的5'-非编码区（5'-NCR）的主要部分存在于健康的抗HCV阴性个体的外周血单核细胞（PBMC）的DNA部分中经过测试。此外，我们设计了实验来证明这些发现的特异性，方法是使用限制酶对PCR之前的靶DNA（PCR之前的酶切）和PCR之后的产物（PCR后的酶切）进行酶切测定。总之，我们的研究表明，HCV内部核糖体进入位点（IRES）结构的主要部分至少包含在测试个体的DNA中。

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