酶联免疫发光法检测金葡菌肠毒素(SE)B和SEC1方法的建立  

Establishment of chemiluminescence immunoassay for detecting staphylococcal enterotoxin B and C1

作　　者：陈立杰[1] 刘振世[2] 董邦权[1] 杨琨[1] 李琦[1] 金伯泉[1] 

机构地区：[1]第四军医大学免疫学教研室,陕西西安710032 [2]北京倍爱康生物技术股份有限公司,北京100070

出　　处：《细胞与分子免疫学杂志》2006年第5期668-669,673,共3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(O5XJZ006)

摘　　要：目的:比较酶联免疫测定法(ELISA)和酶联免疫化学发光法(CLIA)在检测SEB、SEC1中的敏感性及稳定性。方法:常规法制备、纯化抗SEB和SEC1单克隆抗体(mAb),以碱性磷酸酶(AP)标记的FMMUSEB.D6和FMMUSEC1.C4mAb为检测抗体,FMMUSEB.B4和FMMUSEC1.G8mAb为包被抗体,以单磷酸酚酞(PMP)为显色底物,以LumigenAPS5为发光底物。结果:成功地建立了敏感、特异检测SEB、SEC1的ELISA和CLIA方法,CLIA较传统ELISA具有灵敏度高、线性范围宽和省时等优点。结论:CLIA法是一种比传统ELISA更优越的免疫学检测方法,在可疑SE污染标本检测、流行病学调查等领域具有良好的应用前景。AIM： To establish chemiluminescence immunoassay （CLIA） for detecting staphylococcal enterotoxin B （SEB） and C1 （SEC1） and compare its sensitivity and stability with ELISA. METHODS： The anti-SEB and SEC1 monoclonal antibodies （mAb） were purified by Q Sepharose Fast Flow chromatographic column. The alkaline phos- phatase （AP） conjugated mAbs FMMU-SEB. D6 and FM- MU-SECI. C4 were used as detecting antibodies and the FMMU-SEB. B4 and FMMU-SEC1. G8 mAbs were used as coating antibodies in both methods. Phenolphthalein mono- phosphate （PMP） and lumigen APS-5 were employed as substrates for AP in ELISA and CLIA, respectively. The light was detected and measured by the GENios analyzer （TECAN Group Ltd. ）. The sensitivities and detect ranges of CLIA and ELISA methods were compared. RESULTS. Compared with ELISA, CLIA was more sensitive （0. 1 ng/mL vs 0. 39 ng/mL） and timesaving. Furthermore, the liner range of CLIA was broader than that of ELISA （0. 78 - 50 ng/mL vs 3. 125-50 ng/mL）. CONCLUSION： CLIA for detecting SEB and SEC1 are established successfully which may be useful in food monitoring, epidemiology survey and detecting SE contaminated samples in environment.

关 键 词：酶联免疫测定(ELISA) 酶联免疫化学发光测定(CLIA) 金黄色葡萄球菌肠毒素(SE) 检测 

分 类 号：R392.11[医药卫生—免疫学]