山东地区输血传播病毒(TTV)检测及部分基因克隆和序列测定

目的 了解TTV山东分离株感染状况及基因变异的情况。方法应用逆转录聚合酶链反应(PCR)检测26例山东地区非甲-庚型肝炎和12例肝细胞癌患者血清中TTV DNA,并对阳性扩增的产物利用PCR技术直接克隆和测序,分析其基因变异的情况。结果26例非甲-庚型肝炎中11例TTV DNA阳性(42％)。对其中两株(TTVSD4、TTVSD5)部分基因克隆测序,并与日本株(ABOO8394)相比较,其核苷酸旬同源性为99.9％和100％。而12例肝细胞癌患者中3例TTV DNA阳性(25％),对其中一株(TTVSD6)部分基因克隆与测序,与日本株(ABOO8394)相比较,其核苷酸序列同源性为99％;TTV山东株三株间核苷核同源性均为99％。结论本研究证实山东地区非甲-庚型肝炎和肝细胞癌患者中存在着较高TTV感染,TTV感染可能具有嗜肝性,而且可能与肝功能损害有关,是引起非甲-庚型肝炎的重要病原。

Detection of transfusion transmitted virus(TTV) infection and its partial gene cloning and sequencing in Shandong

Objective To investigate the prevalence of TTV infection and genetic variation of TTV isolated in Shandong Province. METHODS TTV DNA were amplified and detected by PCR from sera of patients with NonA - G hepatitis and hepatocellular carcinoma. The PCR products was cloned and sequenced. RESULTS TTV DNA was positive in 11 of 26 patients(42.3%)with nonA - G hepatitis. Partial gene of two TTV isolates(TTVSD4, TTVSD5) was cloned, sequenced and compared with known sequence of TTV isolates in Japan (ABOO8394) reported by Okamoto.The nucleotide homology were 99% and 100% .TTV DNA was positive in 3 of 12 patients(25%) with hepatocellular careinoma. Partial gene of TTV isolates (TTVSD6) was cloned, sequenced and compared with known sequence of TTV isolated in Japan( ABOO8394). The nucleotide homology was 99 % . CONCLUSION The TTV infection rate is high in Shandong. TTV might be hepatotropic, maybe correlated with the impariment of liver function. So, it is likely to be the pathogen of nmA - G hepatitis.