新型凝集技术快速检测产肠毒素性大肠杆菌菌毛的研究  

Research of new agglutination test for rapid detection of ETEC fimbriae

作　　者：李星月[1] 董秀梅[1,2] 陶婧 李婷婷[1] 张萍[1] 冯博[3] 师东方 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030 [2]农业部动物疫病病原生物学重点实验室东北科学观测实验站,黑龙江哈尔滨150030 [3]绥化市动物疫病预防与控制中心,黑龙江绥化152000

出　　处：《中国预防兽医学报》2018年第1期29-34,共6页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家科技支撑计划项目(2012BAD12B03;2012BAD12B05)

摘　　要：为建立以彩色二氧化硅微球(CSN)为载体,快速检测产肠毒素性大肠杆菌(ETEC)菌毛K88ab、K99、987p和F41的新型凝集技术,本研究采用反相微乳液法制备二氧化硅微球,将其与菌毛单因子血清偶联制成免疫彩色二氧化硅微球(ICSN),通过优化反应条件建立了快速检测ETEC菌毛的方法。特异性试验结果显示,ICSN只与携带其对应菌毛的菌株发生反应,与携带其它菌毛的菌株无交叉反应;敏感性试验结果显示菌液的最小检出浓度为1.0×10~6cfu/m L^5.1×10~6cfu/m L;稳定性试验显示4℃保存30 d,ICSN特异性、敏感性无明显变化;双盲样品检测准确率为100%。本研究有助于ETEC菌毛的快速检测和幼畜大肠杆菌性腹泻的诊断。To establish a new agglutination technique for detection of Enterotoxigenic Escherichia coli （ETEC） pili K88ab, K99, 987P and F41 genes based on colored silica microspheres （CSN） synthesized by reverse microemulsion method. The CSN surface was modificated by fimbriae mono-specific serum to prepare immune colored silica microspheres （ICSN）. Then the method for rapid detection of ETEC pilus was established by optimization of reaction conditions. The results showed that the prepared ICSN was only reacted with the corresponding strains and there was no cross reaction with the other strains, and the detection limit was 1.0×106 cfu/mL-5.1×106 cfu/mL. The specificity and sensitivity of ICSN preserved at 4 ℃ for 30 days had no obvious change. The accuracy rate of the detection for double blind samples was 100%. This research established an effective method for detection of ETEC fimbriae and the diagnosis of piglets and calves diarrhea caused by ETEC.

关 键 词：产肠毒素性大肠杆菌 菌毛 单因子血清 二氧化硅微球 新型凝集技术 

分 类 号：S852.4[农业科学—基础兽医学]