猪附红细胞体双抗夹心ELISA检测方法的建立  

Establishment of Sandwich ELISA for Detecting Mycoplasma suis

作　　者：朱凝瑜[1] 袁聪俐[1] 杨志彪[1] 华修国[1] 

机构地区：[1]上海交通大学农业与生物学院,上海200240

出　　处：《上海交通大学学报（农业科学版）》2010年第2期97-101,共5页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：国家863计划(2006AA02Z454);国家自然科学基因面上项目(30871879)

摘　　要：实验通过纯化猪附红细胞体后免疫小鼠及家兔制备阳性血清,建立了猪附红细胞体双抗夹心ELISA检测方法。实验结果表明,以鼠阳性血清1:200稀释后作为捕获抗体,4℃包被过夜,1%明胶溶液37℃封闭1h,兔阳性血清1:400稀释后作为检测抗体,辣根过氧化物酶标羊抗兔IgG抗体1:8000稀释,37℃显色10min为最佳反应条件。同时进行特异性、敏感性、重复性实验。该方法对猪附红细胞体最低检出量为3.812μg·mL-1,具有良好的特异性、敏感性和重复性,有望作为群体检测方法。In the present study, a double-antibody sandwich enzyme-linked immunosorbent assay (sandwich ELISA) for Mycoplasma suis detection was established. The Mycoplasma suis antibody was isolated from experimental infected Kunming mouse and New Zealand rabbit with purified Mycoplasma suis antigen. Mouse original antibody was diluted with 200 fold as the capture antibody and antibody of rabbit diluted 400 as the detection antibody. Reaction condition was optimized with that coating in 4 ℃ overnight, blocking in 37 ...

关 键 词：附红细胞体 双抗夹心ELISA 检测 

分 类 号：S858.28[农业科学—临床兽医学]