TTV病毒检测方法的建立及在维吾尔族人群中的应用

Development and preliminary application of a PCR method for detection of TTV DNA

目的 建立TTV-DNA聚合酶链反应方法并应用于新疆地区不同人群TTV感染的检测。方法 根据已报道的TTV基因序列(DDBJ序列号:AB008394),通过引物设计软件SQNCE和OLIGO在其ORF_1区设计一对PCR引物,扩增产生一个315bp的扩增片段,产物经Bg1Ⅱ酶切分别产生一个219bp和96bp的酶切片段。结果 用建立的PCR方法检测临床标本,在15例维吾尔族转氨酶升高的非甲～非庚型肝炎患者血清标本中检测到6份TTV DNA阳性,表明新疆地区存在TTV感染,维吾尔族人群中有较高的TTV感染率。结论 我们建立的TTV DNA-PCR灵敏度高、特异性好,可用于各类人群TTV-DNA的检测。

Objective To establish a polymerase chain reaction for the detection of TTV-DNA in Xinjiang of different populations and patients with various liver diseases. Methods By using the software of SQNCE and OLIGO, a set of primers was designed based on the reported sequence of TTV (access number AB008394 in DDBJ) . The amplified product was about 315 bp in length and the specificity of the method was confirmed by Bgl II to produce two fragments of 219 bp and 96 bp respectively. Results The PCR method was also successfully applied to clinical samples. TTV DNA was detected in 6 (40% ) of 15 sera of Uigur whose alanine aminotransferase (ALT) were high but no seriological markers of known hepatitis viruses. Conclusion The TTV DNA-PCR has high sensitivity and specificity. It can be used for the detection of TTV DNA in sera of different populations and patients with various liver diseases. The above results suggest that TTV exists in Xingjiang and there was a high incidence of TTV in the Uigur.