输血传播病毒-DNA聚合酶链方法的建立及应用

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

目的 :建立输血传播病毒 (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 :AB0 0 8394) ,通过引物设计软件SQNCE和OLIGO在其ORF1区设计一对PCR引物 ,扩增产生一个 315bp的扩增片段 ,产物经BgⅠⅡ酶切分别产生一个 2 19bp和 96bp的酶切片段。结果 :用建立的PCR方法在 15例维吾尔族转氨酶升高的非甲 -非庚型肝炎血清标本中检测到 6份TTV -DNA阳性 ,表明新疆地区存在TTV感染 ,维吾尔族中有较高的TTV感染率。结论 :我们建立的TTVDNA -PCR方法灵敏度高、特异性好 ,可用于各类人群TTV -DNA的检测。

Objective:[WT5BZ]To establish a new PCR for detection of TTV-DNA among various populations and patients with liver diseases in Xin Jiang.[WT5HZ]Methods:[WT5BZ]Based on the reported sequence of TTV(access number AB008394 in DDBJ),a set of primers was designed by the software of SQNCE and OLIGO,The amplified product was about 315 bp and the specificity of the method was confirmed by BgⅠⅡto produce two fragments of 219 bp and 96 bp respectively.[WT5HZ]Results:[WT5BZ]The PCR method was also successfully applied to clinical samples.TTV DNA was detected in 6 of 15(40%) sera of Uigur whose alanine aminotransferase(ALT) were high but no seriological markers of known hepatitis viruses.[WT5HZ]Conclusion:[WT5BZ]The TTV DNA-PCR has high sensitivity and speciticity.It can be used for the detection of TTV DNA in sera of different populations and patients with various liver diseases.The results suggest that TTV exist in Xingjiang and there is a high incidence of TTV in the Uigur [WT5HZ]