单链构象多态性分析研究HCV NS5A区基因变异

The genomic population of HCV NS5A region using the single-strand conformation polymorphism

用逆转录、套式不对称聚合酶链反应(PCR)制备单链DNA,经凝胶电泳,Southern转移后与地戈辛标记探针杂交.根据单链DNA条带数目对52例慢性肝病患者血清进行HCV NS5A区基因变异程度分析.结果90%的慢迁肝、35%的慢活肝、16%的肝硬化与10%的肝癌患者出现1条单链DNA带,20%的慢活肝、42%的肝硬化、60%的肝癌患者呈现多条单链DNA带,慢迁肝则未见多条单链DNA带.临床参数分析发现单链DNA条带数的多少与肝脏病理改变有关(P<0.05).

The sera from 52 patients with chronic hepatitis C virus infection were analyzed using single-strand conformation polymorphism (SSCP) technique. In the SSCP,an asymmetric PCR was carried out on the 455 bp products of the first round PCR at the NS5A region and the number of band of single srtandof DNA which was reacted with cDNA probe specific for the NS5A region after gel electrophoresis was analyzed. In chronic persistent hepatitis,the band of single strand of DNA was limited to one;however,two or more bands of DNA were frequently detected in chronic active hepatitis and liver cirrhosis. In about half of patients with hepatocellular carcinoma,3 or more bands were found. The number of bands tended to increase in parallel with the progression of liver diseases. The multivariate analysis showed that the progression of liver diseases was the independent factor of viral diversity (P<0. 05)