柯萨奇B3病毒VP1基因在原核中的表达及其临床意义.pdfVIP

毕胜利等 柯萨奇B3病毒VP1基因在原核中的表达及其临床意 箜 · 847· 柯萨奇B3病毒VP1基因在原核中的表达及其临床意义 毕胜利 曲 萌 陈廷友 宋玉国 (北华大学医学院，吉林132013) 中国图书分类号 11373．23 文献标识码 A 文章编号 1000-484X(2007)09-0847-05 [摘 要] 目的：研究柯萨奇B3病毒(CVB3)VP1基因在大肠杆菌中的表达及其临床意义。方法：将柯萨奇B3病毒中 国分离株VP1基因克隆入PQE．30载体，转导人大肠杆菌(M15)中，使CVB3的VP1基因在大肠杆菌中得到稳定的表达，采用 NAT亲和方法纯化，间接ELISA方法做免疫学活性鉴定。结果：表达的VP1产物与抗柯萨奇B3病毒的兔抗CVB3(多克隆抗 体)血清产生较强的免疫反应，与天然的CVB3蛋白抗原(病毒组织培养抗原)的抗原性近似。应用无关兔抗体对照呈阴性反 应。应用表达的VP1产物作为抗原，对临床急性病毒性心肌炎患者血清进行了特异性IgM酶联免疫吸附试验检测，其结果与 组织培养的CVB3制备的细胞蛋白抗原对上述患者血清的特异性IgM检测结果基本一致。结论：采用基因工程方法制备出的 CVB3．VP1蛋白抗原产量高，纯化后免疫反应原性基本没有变化。试用表达CVB3的VP1基因工程抗原代替目前实验用有感 染危险的病毒培养抗原的方法，快速、特异地检出CVB3的感染，为急性心肌炎的早期诊断和及时的临床治疗提供可靠的实验 室诊断指标。 [关键词] 柯萨奇B3病毒(CVB3)；VP1基因；基因表达 Expression of coxsackievirus group B3 gene fragment encoding VP1 in procaryon and clinical significance BI Sheng—Li，QC Meng，CHEN Ting—You，SONG Yu—Guo．Jilin Medical College of Beihua University，Jilin 132013， China [Abstract] Objective：To study the expression of coxsackievirus group B3 gene fragment encoding VP1 in procaryon and to ex— plore its application．Methods：Stablie expression of VP1 gene of CVB3 in E coli was obtained．The expressed protein was purified by NAT chromatography and its immunoactivity was identified by indirect ELISA．Results：The expressed product of VP1，similar to native protein antigen of CVB3，could strong bind with the mouse antibody senlm against CVB3(polyclonal antibody)．Irrelevant monoclonal antibody as contrast presents negative activity．Using the expressed VP1 product，we have had a special lgM EL1SA for the patient~ser- am of the clinical acute viral myocarditis．The result was saine with the cellular protein antigens of the tissue—cultivated CVB3．Conclu- sion：The protein antigen CVB3一VP1 which is obtained by the method of gene engineering has character of high product，and its immu— noactivity after being purified was basically unchanged．At present，this kind of antigen is diffic