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目的分析中国丙型肝炎病人ＨＣＶ基因组３′端非编码区（３′ＮＣＲ），以促进对ＨＣＶ基因组复制机制的研究。方法采用两种方法，从上海地区感染ＨＣＶ的病人血清中，扩增获得ＨＣＶ基因组３′端非编码区：一是用套式ＰＣＲ直接扩增，二是先分别获得ＨＣＶ３′ＮＣＲ的前半部分和后半部分，再将两片段进行融合ＰＣＲ。ＰＣＲ产物进行测序后作同源性分析。在此基础上，建立了针对３′端非编码区的ＲＴＰＣＲ方法，并与基于５′端非编码区的ＲＴＰＣＲ方法检测ＨＣＶＲＮＡ的特异性和灵敏度比较。结果序列分析表明，中国丙型肝炎病人ＨＣＶ基因组３′非编码区由４部分组成：高度变异区、Ｐｏｌｙ（Ｕ）区、Ｐｏｌｙ（Ｕ／Ｃ）区和９８碱基区。同源性分析显示，９８碱基区在不同分离株间高度保守并与国外报道株一致，而Ｐｏｌｙ（ＵＵＣ）区存在较大差异。３′端非编码区和５′非编码区ＲＴＰＣＲ检测血清ＨＣＶＲＮＡ有较高符合率（９５％）。结论ＨＣＶ基因组３′端非编码区的３′末端（９８碱基），在不同分离株间的高度保守性提示，该区在ＨＣＶ基因复制中起重要作用。基于３′非编码区的ＲＴＰＣＲ方法，将有助于ＨＣＶ感染的诊断。
Objective To analyze the 3 ’noncoding region (3’NCR) of HCV genome in patients with hepatitis C in China so as to promote the study of the mechanism of HCV genome replication. Methods Two methods were used to amplify the 3 ’untranslated region of HCV genome from the serum of patients infected with HCV in Shanghai. The first was amplified by nested PCR and the other was the first half of HCV 3’ The second half, then two fragments for fusion PCR. PCR products were sequenced for homology analysis. On this basis, RT-PCR method for 3 ’non-coding region was established and compared with RT-PCR method based on 5’ non-coding region to detect the specificity and sensitivity of HCVRNA. Results The sequence analysis showed that the 3 ’non-coding region of HCV genome of HCV patients in China consisted of 4 parts: highly variable region, Poly (U) region, Poly (U / C) region and 98 base region. Homology analysis showed that the 98 base region was highly conserved among different isolates and was consistent with the reported foreign strains, while there was a big difference in the Poly (UUC) region. The 3 ’non-coding region and 5’ non-coding region RT PCR detection of serum HCVRNA have a high coincidence rate (95%). Conclusion The 3 ’end (98 bp) of the 3’ untranslated region of HCV genome is highly conserved among different isolates suggesting that this region plays an important role in HCV gene duplication. The RT-PCR method based on the 3 ’non-coding region will be helpful for the diagnosis of HCV infection.