酵母表达的风疹病毒E1在ELISA检测中应用

目的将毕赤酵母表达系统中表达的E1糖蛋白作为包被抗原,建立一种优于病毒作为包被抗原的风疹病毒(rubellavirus,RV)抗体ELISA检测方法。方法将重组包膜糖蛋白(E1)抗原和风疹病毒分别作为包被抗原,用间接ELISA法检测90份临床血清标本,同时用晶美(JINGMEI)进口分装试剂盒和德国RECI公司试剂盒进行检测;将RECI试剂盒作为金标准,计算另外三者检测结果的特异性、灵敏性和符合率;比较他们之间的差异有无统计学意义,特别是重组抗原与病毒抗原作为包被抗原检出RV抗体阳性率的差异有无统计学意义。结果重组蛋白重复性实验结果经统计学处理,P>0·05,差异无统计学意义;RECI试剂盒与重组抗原、病毒抗原的检测结果差异有统计学意义(P<0·05);重组抗原与病毒抗原的检测结果差异有统计学意义(P<0·05),作为包被抗原,重组抗原优于病毒抗原。结论用毕赤酵母表达的RVE1重组蛋白作为包被抗原进行ELISA检测优于病毒作为包被抗原,具有广阔的应用前景。

Application of recombinant glycoprotein E1 of rubella virus in ELISA

Objective To establish a specific and sensitive serological method,ELISA,for detection of rubella virus(RV)antibody using yeast expression products as coating antigen and to improve its sensitivity and specificity.Methods The yeast expression products and RV culture medium were used as coating antigens respectively to assay 90 sera.The same sera were also assayed by JINGMEI kit and Germany RECI kit.The results of Germany kit,as a golden standard,were compared with those of the other three methods in specificity,sensitivity and accordance rate.And the statistical test was used to compare the differences among these methods,and also to evaluate which was more useful for coating antigen,recombinant antigen or RV culture medium.Results The results of repeated experiment of recombinant protein showed that there was no statistical significance between two experiments(P>0.05).The difference of the two positive rates between RECI kit and recombinant antigen was significant(P<0.05).The difference of the two positive rates between RECI kit and RV culture medium was significant(P<0.05).There was statistical significance between those of recombinant antigen and RV culture medium(P<0.05).Conclusion As coating antigen,the recombinant E1 Protein is better than that of RV culture medium in ELISA method for detection of RV antibody.