河南TT病毒全基因序列测定及TTV感染检测方法的研究

目的 :为了分析河南省TTV的基因结构、特征及基因变异 ,建立适合我国人群的诊断方法。方法 :采用巢式聚合酶链式反应 (PCR)技术从河南省血液中心献血员TTV阳性血清中扩增TTV全基因 ;用PCR方法扩增ORF2片段 ,进行原核表达 ,并转化大肠杆菌 ,进行诱导表达。用SDS PAGE分析表达产物。结果 :在国内较早地克隆测定了一株TTV河南分离株的基因序列 ;建立了检测TTV感染的PCR检测方法 ;在国内较早地表达出TTV重组蛋白抗原 ,抗原具有良好的活性。结论 :用表达的抗原初步建立了检测血清抗 TTVIgG抗体的间接ELISA方法 ,可有效地应用于TTV感染的检测。

Sequencing of TTV gene from a Henan isolate and development of diagnostic methods for detection of TTV infection

Objective: This study aimed to clone and sequence the complete TTV gene from a Henan isolate,and to develop the diagnostic methods for detection of TTV infection.Methods: Using PCR to sequence the TTV gene and clone to T-vector.The recombinant plasmid was transformed into E.coli strain M15 and induced with IPTG.We purified the specific expressed protein through Ni-NTA column,the purification was analyzed by Western Blotting.Results: Cloning and sequencing the TTV gene from Henan isolate.The TTV recombinant protein antigen was expressed in E.coli.Conclusion: The polymerase chain reaction are useful in diagnosis of TTV infection .The recombinant protein antigen can used for diagnosis of TTV infection.