摘要
目的构建表达风疹病毒蛋白抗原的重组质粒及工程菌,获得纯化的E1蛋白抗原,用于检测风疹病毒特异性抗体IgM。方法克隆表达风疹病毒E1重组蛋白,利用产物建立IgM捕获ELISA方法鉴定其抗原性及实用性。结果表达纯化的E1蛋白经酶标记建立IgM捕获ELISA方法,检测53份抗风疹病毒IgM阳性血清和67份阴性血清,用酶标记E1蛋白建立的捕获ELISA法阳性检出率98.1%,阴性检出率100%,与意大利SORIN公司试剂盒检测结果比较,无统计学意义(P>0.05);其中1份风疹病毒(IgM)阳性血清1∶16稀释后仍能与抗原反应,初步表明E1蛋白抗原表位有较好的抗原特异性。结论高效表达纯化的E1蛋白抗原性强,利用其建立的IgM捕获ELISA方法,可用于检测风疹病毒抗体。
Objective To canstruct the recombinant plasmid and engineering bacteria which express Rubella Virus protein antigen, El protein antigen can be producted and purified. This products can be used to detect RV specific antibody IgM.Methods Recombinant protein of Rubella Virus E1 was Cloned and expressed to establish IgM capture ELISA for indentifying its anfigenicity and practicality.Results Through IgM capture ELISA to detect 53 RV-positive sera and 67 RV-negative sera, the positive rate was 98.1% and negative rate was 100%. Compared with SOR/N company, there is no significant differemce（ P 〉 0.05）.One of the RV- positive sera can respongse to antigen after 1 ： 16 dilution,and this indicate initially E1 protein epitope had better spacificity. Conclusion IgM capture ELISA estabilshed by E1 protein which expressed and purified with high performance and antigencity can be used to detect RV antibody.
出处
《中国实验诊断学》
北大核心
2009年第4期433-435,共3页
Chinese Journal of Laboratory Diagnosis
基金
吉林省科技厅课题(课题编号200705204)
