小反刍兽疫病毒N蛋白B细胞表位预测、合成及间接ELISA方法的建立  

Prediction of B Cell Epitopes,Synthesis And Development of Indirect ELISA for N Protein of Peste Des Petits Ruminants Virus

作　　者：曾少灵[1] 阮周曦[1] 唐金明[1] 廖立珊[1] 孙洁[1] 林庆燕[1] 吕建强[1] 叶奕优[1] 祝贺[2] 李希强 王红英[1] 杨俊兴[1] 花群义[1] 

机构地区：[1]深圳出入境检验检疫局动植物检验检疫技术中心,广东深圳518045 [2]云南出入境检验检疫局检验检疫技术中心,云南昆明650028 [3]内蒙古牙克石市动物疫病防控中心,内蒙古牙克石022150

出　　处：《动物医学进展》2015年第8期40-44,共5页Progress In Veterinary Medicine

基　　金："十二五"国家高技术研究发展计划项目(863计划)(2012AA101301);"十二五"国家科技支撑计划项目(2013BAD12B03);深圳检疫局科研项目(SZ2014203);国家质检总局科研项目(2015IK244)

摘　　要：基于氨基酸序列和采用Gamier-Robson和Chou-Fasman等多种方法分析的α-螺旋、β-折叠和转角等二级结构结果,预测小反刍兽疫病毒N蛋白的B细胞抗原表位。设计4段多肽并进行人工合成,作为包被抗原建立小反刍兽疫血清抗体间接ELISA检测方法,用于检测小反刍兽疫山羊临床血清样品,以对预测的B细胞抗原表位进行验证。结果显示,研制的间接ELISA敏感性为96.0%,特异性为96.25%,与进口标准竞争ELISA试剂盒的符合率为96.15%。结果表明,研制的间接ELISA方法适用于小反刍兽疫临床血清抗体的检测。Based on the amino acid sequence and the analysis results of secondary structures such as α-he-lix,β-sheet and turn regions,etc,the B cell epitopes for N protein of peste des petits ruminants virus were predicted by methods of Gamier-Robson and Chou-Fasman.Four artificial peptides were designed,synthe-sized and used as coating antigens,by which the indirect ELISA of serum antibody test for peste des petits ruminants virus was developed.The comparison test on goat clinical serum samples was carried out.Com-pared with the imported standard competitive ELISA,the sensitivity and specificity of the developed indi-rect ELISA were 96.0% and 96.25%,and the coincidence rate of the two methods was 96.15%.So the developed indirect ELISA method coating with synthetic peptides is suitable for clinical application.

关 键 词：小反刍兽疫病毒 N蛋白 B细胞表位 酶联免疫吸附试验 

分 类 号：S852.659.5[农业科学—基础兽医学]