ELISA法检测非洲猪瘟抗体实验条件的优化  

Optimizing the Conditions of an Indirect ELISA for Detection of African Swine Fever Virus Antibody

作　　者：张永强[1] 任伟杰[1] 吴晓东[1] 王筱真 李金明[1] 王志亮[1] 

机构地区：[1]中国动物卫生与流行病学中心/外来动物疫病研究中心,山东青岛266032

出　　处：《中国动物检疫》2014年第11期97-100,共4页China Animal Health Inspection

基　　金：公益性行业(农业)科研专项经费(201103008)

摘　　要：本文以灭活非洲猪瘟病毒为包被抗原,分析了包被浓度、酶标二抗、显色底物对ELISA法检测非洲猪瘟抗体的影响,以期获得最适反应条件。以2个阳性血清和一个阴性血清为待检样品,用不同包被浓度、2种酶标二抗和2种显色底物进行ELISA反应,记录吸光值,分析阴阳性样品吸光值差异,优化阴性样品背景值,计算P/N值。结果表明,包被浓度为15μg/m L、二抗为HRP标记蛋白A、显色底物为OPD时,两个阳性样品P/N值分别为4.920和7.259,为本方法的最适反应条件。In the study,the effects of antigen coating concentration,enzyme labeled antibody and chromogenic sub-strate in an ELISA with inactivated African swine fever virus as coating antigen were analyzed for detection of ASFV antibody. In order to obtain the optimum reaction conditions,different coating antigen concentrations,2 type enzyme-labeled antibody and 2 type chromogenic substrates were used in the ELISA to detect 2 positive serum samples and 1 negative serum sample. The result showed that the optimum reaction conditions were：the antigen coating concentration was 15μg/mL;Protein A-HRP was used as labeled antibody,and OPD as chromogenic substrate and the P/N value of 2 positive serum samples were 4.920 and 7.259.

关 键 词：ELISA 非洲猪瘟抗体 试验验条件 优化 

分 类 号：S858-28[农业科学—临床兽医学] S852.659.1[农业科学—兽医学]