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目的对TTV阳性扩增产物进行克隆及测序,以了解西安地区TTV基因型及基因结构的特点。方法采用巢式PCR从转氨酶活性异常增高的患者血清中,得到TTV阳性扩增产物,将其插入pGEM-Teasy载体,进行克隆及序列分析。结果获得1个pGEM-TTV重组载体。序列分析表明,插入片段为196bp。该序列与AF065400株等对应位置的核苷酸同源性分别为:AF065400中国株94.9%、AF351132中国株98.0%和NC-002076日本株99.0%。结论西安地区TTV阳性扩增序列与报道的AF065400株、AF351132株和NC-002076株的序列具有高度的同源性,属同个基因型别。
Objective To clone and sequence TTV positive amplification products to understand the characteristics of TTV genotype and gene structure in Xi’an. Methods Nested PCR was used to amplify TTV-positive products from the sera of patients with abnormally elevated transaminase activity. The product was inserted into pGEM-Teasy vector and cloned and sequenced. Results One pGEM-TTV recombinant vector was obtained. Sequence analysis showed that the insert was 196 bp. The nucleotide homology of this sequence to the corresponding position of AF065400 and so on were 94.9% for AF065400, 98.0% for AF351132 and 99.0% for NC-002076 respectively. Conclusion The sequence of TTV positive amplification in Xi’an region is highly homologous with the reported sequences of AF065400, AF351132 and NC-002076, which belong to the same genotype.