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目的分析ＴＴＶ（Ｔｒａｎｓｆｕｓｉｏｎｔｒａｎｓｍｉｔｅｄｖｉｒｕｓ）深圳分离株ＯＲＦ１部分基因序列。方法在ＴＴＶＯＲＦ１设计引物，建立巢式聚合酶链反应（Ｎｅｓｔｅｄ－ＰＣＲ），检测４０例广东地区非甲－庚型肝炎患者血清中ＴＴＶＤＮＡ。对ＰＣＲ产物进行分子克隆，以荧光法（ＡｐｐｌｉｅｄＢｉｏｓｙｓｔｅｍｓ，３７３Ａ）测序。结果４０例非甲－庚型肝炎中２１例ＴＴＶＤＮＡ阳性（５２．５％）。对其中一株ＴＴＶ（ＳＺ１）ＯＲＦ１部分基因克隆、测序，并与Ｏｋａｍｏｔｏ等报道的２株（Ｎ２２、Ｇ１ａ）相比较，其核苷酸序列的同源性分别为９６．７％与９７．４％。结论本研究证实我国华南地区存在ＴＴＶ感染。ＴＴＶ的分子克隆与测序及ＰＣＲ诊断技术的建立对国内进一步开展ＴＴＶ感染的诊断与流行病学调查均具有重要意义
Objective To analyze the partial ORF1 gene sequence of Shenzhen isolate of TTV (Transfusiontransmited virus). Methods Primers were designed in TTVORF1 to establish a nested polymerase chain reaction (Nested-PCR) to detect the serum TTVDNA in 40 patients with non-A-G hepatitis in Guangdong Province. The PCR products were molecularly cloned and sequenced by fluorescence (Applied Biosystems, 373A). Results TTV DNA was positive in 21 (52.5%) of the 40 non-A-G hepatitis patients. One part of TTV (SZ1) ORF1 gene was cloned, sequenced and compared with the two strains reported by Okamoto et al (N22, G1a), the nucleotide sequence homologies were 96.7% and 97.4 %. Conclusion This study confirms the existence of TTV infection in southern China. The molecular cloning and sequencing of TTV and the establishment of PCR diagnosis are of great significance for the further diagnosis and epidemiological investigation of TTV infection in China