丙型肝炎病毒5’非编码区PCR产物单链构象多态性分析及序列测定

目的：研究慢性肝炎病人血清、肝细胞肝癌组织中丙型肝炎病毒（ＨＣＶ）５＇非编码区变异情况．方法：收集１０例慢性肝炎病人及４例肝细胞肝癌病人血清和肝细胞肝癌组织，检测抗ＨＣＶ抗体阳性，进行ＨＣＶ５＇非编码区ＲＮＡ的逆转录ＰＣＲ扩增，ＰＣＲ产物进行单链构象多态性分析，并对部分病例序列进行测定．结果：６例慢性肝炎病人血清及３例肝细胞肝癌组织中检测到ＨＣＶＲＮＡ，单链构象多态性分析显示各例条带之间无明显差异，对１例肝细胞肝癌组织中ＨＣＶ５＇非编码区ｃＤＮＡＰＣＲ产物的序列测定表明它与ＨＣＶ－１的同源性为９７．３％，与ＨＣＶ－ＢＫ的同源性为９７．７％．结论：我们的结果提示在西安地区ＨＣＶ５＇非编码区几乎是一样的，并进一步证实不同地区ＨＣＶ５＇非编码区的高度保守性．

The single strand conformation polymorphism analysis and sequencing of the PCR products of HCV 5'''' untranslated region

Objective: To study the heterogeneity of 5' untranslated region (5' UTR) of hepatitis C virus (HCV) in chronic hepatitis (CH) and hepatocellular carcinoma (HCC). Methods: The serum of 10 patients with CH and the serum and tissues of 4 patients with HCC were collected and confirmed anti HCV antibody positive. 5' UTR RNA were detected by RT PCR for single strand conformation polymorphism (SSCP). PCR products (257 bp) were denatured into single strand and analyzed by electrophoresis on a 8% polyacrylamide gel, and determined the number of bands. The PCR product of one case of HCC was cloned into pUC19 plasmid and its sequence was analyzed by ABI370 automated DNA sequence analysis machine. Results: HCV 5' UTR RNA was detected in 6 cases of CH and 3 cases of HCC. The SSCP bands of 5' UTR PCR product showed no obvious difference between cases. Sequence analysis of HCV 5' UTR in one case of HCC showed that it shared 97.3% homology with HCV 1 and 97.7% homology with HCV BK. Conclusion: The results indicated that HCV 5' UTR in different cases of our area was almost the same, and further confirmed that the HCV 5' UTR was highly conserved in different countries.