羊驼副结核病间接ELISA检测方法的建立  

Establishment of indirect ELISA for detection of antibody to alpaca paratuberculosis

作　　者：唐泰山[1] 廉慧锋 刘倩[3] 王凯民[1] 祝长青[1] 张常印[1] 雷治海[3] 

机构地区：[1]江苏出入境检验检疫局,江苏南京210001 [2]山西出入境检验检疫局,山西太原030024 [3]南京农业大学动物医学院,江苏南京210095

出　　处：《畜牧与兽医》2013年第11期21-25,共5页Animal Husbandry & Veterinary Medicine

基　　金：国家质检总局科技项目(2008IK03)

摘　　要：利用副结核分支杆菌重组Hed蛋白,建立了检测羊驼副结核分支杆菌抗体的间接ELISA方法。确定了最适抗原包被浓度为3.75μg/mL,最适血清稀释度为1∶50,最佳封闭条件为2%明胶4℃封闭24 h,血清最适作用时间为1.0 h,酶标SPA最适稀释度为1∶5 000,最适作用时间为1.0 h,底物最适作用条件为37℃避光显色10 min。用建立的ELISA方法检测47份羊驼临床血清,显示该方法具有较好的应用前景。The indirect ELISA for detection of antibodies of Mycobacterium paratuberculosis in alpaca was established using recombinant host expression dependent （Hed） protein as coated antigen. The result showed that the optimal concentration of coated antigen was 3.75 μg/ mL, the best sealing condition was under 24 h with 2% gelatin at 4℃, optimal dilution of serum was 1 ： 50, optimal dilution of SPA was 1 ： 5000, and the best substrate reaction condition was at 37℃ for 15 min. Fourty-seven alpaca sera were tested by the indirect ELISA. It in-dicated that the established ELISA is a promising diagnostic method in alpaca.

关 键 词：羊驼 副结核病 重组Hed蛋白 间接ELISA 

分 类 号：S855.1[农业科学—临床兽医学]