戊型肝炎病毒结构区基因的克隆表达及其在诊断中的初步应用

Cloning, sequencing and expressinon of Hepatitis E virus structural gene in E. coli and application of the recombinant products for diagnosis

目的研制戊型肝炎病毒诊断试剂。方法利用融合蛋白表达载体ｐＧＥＸ－３Ｘ表达了戊型肝炎病毒第二读码框区（ＯＲＦ２４０２～６６０）优势抗原表位。表达抗原溶于水，可利用商品化的谷胱甘肽Ｓｅｐｈａｒｏｓｅ－４Ｂ亲合层析柱得到纯化的抗原。结果经应用发现，此基因重组抗原作为酶联免疫试剂的抗原，用于检测血清中抗戊型肝炎病毒抗体，与新加坡进口试剂盒有高度的一致性，和血清中抗体反应性更强。结论预示该基因抗原可能具有较好的反应谱。

The prokaryotic expression vector PEGX-3X was used to express hepatitis E virus (HEV) open reading frame 2(ORF2 402-660).The recombinant protein was soluble in PBS buffer and was purified by the glutathione Sepharose 4B affinity column.When the purified recombinant protein was applied to detect HEV antibodies in sera of clinical patients,the results were quite consistent with the HEV diagnostic kit from Diagnostic Biotechnology Ltd(DBL),Singapore.The recombinant protein was more reactive with HEV antibodies than DBL reagent.