弓形虫SAG1重组蛋白的制备及血清学检测应用  

Expression,purification and characterization of recombinant SAG1 from Toxoplasma gondii

作　　者：陶艳琳[1] 付永锋[2] 赵雪涛[1] 汤宇帆[1] 沈小婷[1] 程训佳[2] 

机构地区：[1]上海市徐汇区疾病预防控制中心,上海200237 [2]复旦大学上海医学院病原生物学系,上海200032

出　　处：《中国卫生检验杂志》2012年第9期2014-2017,共4页Chinese Journal of Health Laboratory Technology

基　　金：上海市公共卫生优秀青年人才培养计划项目(08GWQ020)

摘　　要：目的:以重组弓形虫表面抗原SAG1对上海市徐汇区人群进行弓形虫抗体血清阳性率分析。方法:重组质粒pET-19b-SAG1在大肠杆菌BL21star(DE3)pLysS中诱导表达,经Ni2+亲和层析法纯化目的蛋白后,用ELISA鉴定其抗原特异性。以SAG1重组蛋白为抗原用ELISA方法检测上海市徐汇区人群血清(266份)弓形虫IgG抗体,并与国外试剂盒进行比较。结果:SAG1重组蛋白以包涵体形式存在,产量为0.98 mg/100 ml培养基,纯度达95%以上,并具有较好的抗原特异性。以该重组抗原对上海市徐汇区人群弓形虫血清抗体进行IgG-ELISA检测,显示阳性率为9.4%(25/266)。结论:成功表达提纯了具有较高抗原特异性的SAG1重组蛋白。本次调查上海市徐汇区人群的弓形虫血清抗体阳性率为9.4%,建议加强血清学监测,便于及时采取有效干预措施。Objective：To analyze the situation of Toxoplasma infection among population in Xuhui district of Shanghai city using the recombinant Toxoplasma gondii surface protein SAG1.Methods： Recombinant plasmids pET19b-SAG1 were expressed in E.coli BL21 star（DE3） pLysS.After purification by Ni2＋ gel column chromatography,the antigenicity of recombinant SAG1 was confirmed.The specific IgG antibodies against recombinant SAG1 in the sera of people（266 cases） in Xuhui district were detected by ELISA,and confirmed by foreign diagnostic kit.Results： 0.98 mg recombinant SAG1 with a purity as high as 95% were obtained in 100ml medium and existed in a form of inclusion body.It showed good antigenicity.Serological surveillance of Toxoplasma infection in Xuhui district using the recombinant SAG1 showed positive rate of 9.4%（25/266）.Conclusion： Recombinant SAG1 was expressed successfully with relatively high specificity.The positive rate of anti-toxoplasma IgG in people in Xuhui district of Shanghai city was 9.4%.It was suggested that serological monitoring should be employed,so that effective intervention methods can be used in time.

关 键 词：弓形虫 SAG1重组蛋白 ELISA 

分 类 号：R531.8[医药卫生—内科学]