软海绵酸胶体金免疫检测法的建立与应用  

Construction and application of gold-labeled immunology assay for the detection of okadaic acid

作　　者：蔡春尔[1] 柳俊秀[1] 胡乐琴[1] 贾睿[1] 汪卿[1] 李春霞[1] 何培民[1] 

机构地区：[1]上海海洋大学水产与生命学院,上海201306

出　　处：《国际检验医学杂志》2012年第18期2181-2182,2184,共3页International Journal of Laboratory Medicine

基　　金：海洋赤潮灾害立体监测技术与应用国家海洋局重点实验室开放研究基金资助项目(MATHAB20100309;MATHAB20100205);上海市科委资助项目(08XD14037;10391901900)

摘　　要：目的建立并应用软海绵酸(OA)胶体金免疫层析检测法。方法通过纯化抗OA单抗、制备胶体金、标记单抗、制备胶体金试剂条、样品模拟提取与加标回收检测、贝类染毒提取及检测,建立和优化金标免疫层析法。结果选择30nm胶体金颗粒用于单抗标记,金标单抗浓度为200μg/mL,金标单抗包被量为2.5μL/cm,二抗包被浓度为1.0mg/mL,反应时间为3～5min,对加标贝肉和染毒贝类的检测灵敏度为25ng/mL。结论建立了可用于OA检测的胶体金免疫层析检测法,满足规定的贝类OA含量安全阈值,为腹泻性贝毒检测与食品安全检验提供了新的技术方法。Objective To construct and apply gold labeled immunology assay for the detection of okadaic acid. Methods Gold- labeled immunology testing technology for fast detecting okadaic acid was established and optimized by a series of procedures,inclu ding purification of monoclonal antibody of okadaic acid, preparation of colloidal gold, label of monoclonal antibody, preparation of strip, simulated sample extraction and spiked recovery detection, followed by extraction and testing of exposed shellfish. Results 30 nm colloidal gold particles were used to mark the test strip,the concentration and coated amount of gold labeled monoclonal anti- body were 200 μg/mI, and 2.5 μL/cm, respectively, and the concentration of second antibody was t. 0 mg/mL. The colloidal gold test strip detection in spiked shellfish and exposed shellfish were applied, with detection time of 3--5 minutes and detection limit of less than 25 ng/mL. Conclusion Gold-labeled immunology assay for fast detecting okadaic acid was established,meeting the safe threshold of okadaic acid in shellfish food ruled by many countries and proving technical foundation for detection of okadaic acid in shellfish.

关 键 词：黑软海绵素A 腹泻性贝毒 单克隆抗体 胶体金免疫层析 

分 类 号：R446.6[医药卫生—诊断学]