1型鸭疫里氏杆菌抗体间接ELISA检测方法的建立  

Development and Application of An Indirect ELISA for Detection of Riemerella anatipestifer Serotype 1

作　　者：杨依霏[1] 罗青平[2] 张蓉蓉[2] 艾地云[2] 廖永洪[2] 邵华斌[2] 程国富[1] 

机构地区：[1]华中农业大学动物医学院动物病理实验室,湖北武汉430070 [2]湖北省农业科学院畜牧兽医研究所,湖北武汉430064

出　　处：《动物医学进展》2012年第1期13-18,共6页Progress In Veterinary Medicine

基　　金：公益性行业(农业)科研专项:水禽主要疫病快速诊断与疫苗研制(201003012);湖北省科技厅十一五重大攻关项目:规模化养鸭疫病防控技术集成研究与示范(2009BBB003)

摘　　要：以1型鸭疫里氏杆菌(RA)全基因组保守区域ompA基因的重组表达产物为包被抗原,建立了检测RA血清抗体的间接ELISA方法。原核表达的重组ompA蛋白经纯化后作为包被物,以方阵滴定法确定抗原最佳包被浓度为1.5μg/mL,待检血清最佳稀释度为1∶100。与普通的微量凝集试验相比较,该方法灵敏度高于凝集试验约16倍～128倍。与鸭源大肠埃希菌、鸭源多杀性巴氏杆菌、鸭链球菌、鸭源呼肠病毒、鸭肝炎病毒和鸭瘟病毒感染鸭血清均无交叉反应,表明该方法特异性好。利用该方法检测了雏鸭免疫RA灭活油乳剂疫苗之后血清抗体水平。In this research,an indirect enzyme-linked immunosorbent assay（ELISA） was developed to facilitate early detection of R.anatipestifer infection in ducks.The antigen used was a recombinant 42 ku（ompA） protein,which was the product of the conserved ompA gene in Riemerella anatipestifer type 1 genome.The ompA-based ELISA successfully detected ompA antibody of R.anatipestifer serotype 1.The coated material was recombinant protein ompA after purified.The concentration of antigen was 1.5 μg/mL.Test serum was used after dilution 1∶100.Compared with common micro-agglutination test,ELISA in this research was more sensitive which was 16-128 times higher.There were no cross-reaction with E.coli,P.multocidam,Steptococcus,reovirus,duck hepatitis virus type Ⅰand duck plague virus from ducks,which mean the method was specific.With this method,we test the varition of antibody after injection of inactivated oil emulsion vaccine of RA.

关 键 词：鸭疫里氏杆菌 重组ompA蛋白 ELISA 

分 类 号：S852.617[农业科学—基础兽医学]