摘要
目的:建立检测SEB的双单克隆抗体(mAb)夹心ELISA,并检测在多种基质中SEB的敏感性。方法:制备、纯化抗SEBmAbB4和D6。D6mAb标记辣根过氧化物酶(HRP)作为检测抗体,B4mAb作为包被抗体。结果:成功建立了敏感、特异检测SEB的ELISA方法,检测抗体稀释液和小牛血清中SEB,检测灵感度0.2μg/L,定量线性范围0.78～12.5μg/L,r2=0.992,批间变异系数(CV)<10%,回收率80%～110%。检测50g/L脱脂牛奶、尿液和自来水中的SEB,灵敏度0.39μg/L,定量线性范围0.78～25μg/L,r2=0.997。与SEA和SECl无交叉反应。结论:本方法测量准确,灵敏度高,特异性好,在食品工业和临床标本检测中有广阔的应用前景。
AIM： To establish sandwich ELISA for the detection of Staphylococcal Enterotoxin B （SEB） and characterize the sensitivity and specificity of it in different materials. METHODS： The anti-SEB monoclonal antibodies （mAb） B4 and D6 were employed as capture antibody and detecting antibody respectively after being purified by Q sepharose fast flow chromatography and horseradish peroxidase（HRP） conjugation. RESULTS： The sensitivity of this assay reached 0.2 ng of SEB per mL of PBS with BSA and fetal bovine serum. It reached 0. 39 ng of SEB per mL of 50 g/L skim milk, human urine and water. The concentration curve generated from SEB standard curve was linear with the range of 0.78 - 12.5 μg/L, （ r^ 2 = 0.99 ）. This assay was highly reproducible and the coefficient of variation（CV） was less than 10%. No cross-reactivity between SEA and SECI was found. CONCLUSION： This assay offers a viable method with high sensitivity and specificity for detecting SEB and it may be used for SEB detection in clinical application and food samples.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2007年第8期761-762,共2页
Chinese Journal of Cellular and Molecular Immunology
基金
第四军医大学"211"军事医学研究课题(05XJZ006)
