用套式RT-PCR法检测肝细胞癌中丙型肝炎病毒基因型

目的　检测人肝细胞癌（ＨＣＣ）组织中丙型肝炎病毒（ＨＣＶ）基因型。方法　用ＲＴＰＣＲ法常规检测ＨＣＣ组织和部分患者血清ＨＣＶ，再以巢式ＲＴＰＣＲ法对ＨＣＶ阳性标本中５种ＨＣＶ基因型进行鉴别。对多数标本中ＨＢｓＡｇ和ＨＢＶＤＮＡ也进行了免疫组化和原位杂交检测。结果　ＨＣＣ组织中ＨＣＶ阳性率为５０％（３９／７８），与ＨＢＶ者（６４．４％）差异无显著性意义（Ｐ＞０．０５），两者重叠感染率３４％（２０／５９）。在１０例血清ＨＣＶ阴性ＨＣＣ患者中，３例ＨＣＣ冰冻组织检出ＨＣＶＲＮＡ。在ＨＣＣ组织中共检出３种ＨＣＶ基因型，其中２ａ、１ｂ、３ａ和混合型分别为１７例（４３．６％）、９例（２３．１％）、４例（１０．３％）和２例（５．１％），各基因型间差异有极显著性意义（Ｐ＜０．０１），但２ａ型与１ｂ型间差异无显著性（Ｐ＞０．０５），２ａ型与ＨＢＶ的重叠感染明显较其他ＨＣＶ基因型者常见（Ｐ＜０．０５）。结论　２ａ型ＨＣＶ是ＨＣＣ组织中最常见的基因型并易与ＨＢＶ重叠感染率，ＨＣＣ患者血清ＨＣＶ检测不一定能准确反映ＨＣＣ组织中的ＨＣＶ感染率，其所示基因型也有所不同。

Detection of Hepatitis C Virus Genotypes in Hepatocellular Carcinoma Using RT-Nested PCR

Purpose To detect the hepatitis C virus (HCV) genotypes in human hepatocellular carcinoma (HCC) tissue. Methods HCV in HCC tissue and serum of some HCC patients was routinely detected using RT?PCR method.Then 5 HCV genotypes in HCV positive specimens were identified by nested RT?PCR method.HBsAg and HBV DNA in the most specimens were also examined using immunohistochemistry and in situ hybridization methods. Results The positive rate of HCV in HCC tissue was 50%(39/78),which was not significantly different to that of HBV (64.4%) ( P >0.05).The rate of co infection of HCV and HBV was 34%(20/59).Among 10 HCC patients of whom sera were HCV negative,HCV RNA was found in HCC frozen tissues of 3 cases.3 kinds of HCV genotypes were observed in HCC tissue, including 2a,1b,3a and mixed type with the numberes of 17( 43.6 %),9(23.1%),4(10.3%) and 2(5.1%) respectively.There was high significant difference among all genotypes ( P <0.01),except the difference between type 2a and type 1b( P >0.05).The co infection of type 2a and HBV was most frequent than that of other HCV genotypes( P <0.05). Conclusions Type 2a of HCV was the most common in HCC tissue and prone to co infection with HBV.The detection of HCV in the serum of HCC patients may not reflect the infection rate and genotypes of HCV HCC tissue accurately.