摘要
研究旨在建立基于真核表达的鸭坦布苏病毒(DTV)E蛋白的诊断鸭坦布苏病毒病的间接ELISA方法。利用Bac-to-Bac杆状病毒昆虫表达系统,构建含有DTV E基因的重组杆状病毒。用重组病毒感染昆虫细胞后,SDS-PAGE鉴定出55 ku左右的表达蛋白,Western blot检测该蛋白具有很好的反应原性。以重组Bacmid DTV E蛋白作为包被抗原,建立间接ELISA检测方法。利用该方法对135份来自江苏省的樱桃谷鸭血清样品进行检测,发现其血清阳性率达79.3%。表明建立的间接ELISA检测方法特异性强、重复性好、敏感性高,可以用于鸭坦布苏病毒病的临床诊断。
This experiment was conducted to diagnose duck Tembusu virus （DTV）infection based on the cukaryo expressed DTV E protein through indirect ELISA. The recombinant baculovirus of DTV E gene was constructed by using Bac-to-Bac baculovirus insect expression system. DTV E protein,which efficiently expressed by baeulovirus in Sf9 cells was verified by SDS-PAGE and Western blot. Furthermore, indirect ELISA was developed using recombinant DTV E protein as coating antigen. 135 serum samples of cherry duck from Jiangsu province were detected by this method, the serum positive rate was 79.3%. The results indicated that the indirect ELISA method established in this study had desirable specificity, repeatability as well as sensitivity, and could be used in the diagnosis of DTV infection.
出处
《中国家禽》
北大核心
2016年第1期21-24,共4页
China Poultry
基金
徐州市科技项目(XM13B120)
徐州生物工程职业技术学院科研课题(2013B03)
