狂犬病病毒N、G两种蛋白在间接ELISA方法中的应用比较  

Application and comparison of rabies virus N and G proteins in indirect ELISA

作　　者：王净[1,2] 史利军[2] 李刚[2] 孙全文[1] 吴淑琴[1] 

机构地区：[1]河北北方学院,张家口075131 [2]中国农业科学院北京畜牧兽医研究所,北京100193

出　　处：《中国人兽共患病学报》2013年第6期601-604,共4页Chinese Journal of Zoonoses

基　　金：国家科技支撑计划(No.2013BAD12B05);国际合作项目(No.D32025/17453);国家自然科学基金(No.31172342);国家公益性行业科研专项(No.200903037);河北省畜牧兽医局科技项目(No.2012-3-03)联合资助~~

摘　　要：目的比较狂犬病N、G两种蛋白检测抗体效果。方法本研究先通过比较pET-32a(+)和pGEX-6P-1两种载体表达的狂犬病病毒G蛋白和N蛋白,采用碳酸盐纯化表达量高的两种蛋白,再以纯化的抗原用于检测93份免疫犬血清。结果 (1)以pGEX-6P-1为载体的N、G两种蛋白表达量较高。(2)碳酸盐包被液可以得到纯度较高的蛋白。(3)间接ELISA检测93份抗体,其中56份G蛋白高于N蛋白,37份N蛋白高于G蛋白。(4)血清样本检测结果都为阳性,表明免疫效果良好。结论针对N、G两种蛋白的抗体在动物体内出现时间不同,哪种抗体出现的早,有待于我们进一步研究。The aim of this study is to evaluate the effect of detection for Rabies virus antibody level with recombinant G and N proteins. After comparing of the expression quantity of Rabies virus G and N proteins in pET-32a （＋） and pGEX-6P-1 expression vectors, we chose the recombinant proteins with high expression quantity to purify with the carbonate coating buffer （pH=9.6）. The Rabies virus antibody level in the 93 serums samples were detected by the indirect ELISA using purified pGEX-G and pGEX-N protein as coating antigen respectively. The results showed that the recombinant proteins pGEX-G and pGEX-N with high expression quantity were highly purified with the carbonate coating buffer. The result of the indirect ELISA were positive for all the serum samples, indicated that all of the tested dogs produced antibodies after vaccination. But there are some differences, the detection value of ELISA with pGEX-G were higher than pGEX N in 56 serum samples, for other serum samples, we get the opposite result. The results indicated that anti-G antibody and anti-N antibody were produced at different times in the immune canines. Which kind of antibody appear more early, remains to be further studied.

关 键 词：狂犬病 N蛋白 G蛋白 pET-32a(+) pGEX-6P-1 蛋白纯化 抗体检测 

分 类 号：R373.9[医药卫生—病原生物学]