抗腹泻性贝毒软海绵酸胶体金免疫快速检测技术研究  

A Study on Colloidal Gold Immunoassay Technology for Monoclonal Antibody of Okadaic Acid

作　　者：胡乐琴[1] 马晓康[1] 吴春燕[1] 

机构地区：[1]上海海洋大学水产与生命学院,上海201306

出　　处：《江西农业大学学报》2015年第5期909-913,共5页Acta Agriculturae Universitatis Jiangxiensis

基　　金：上海市教育委员会科研创新项目(14YZ122);科促会联盟项目

摘　　要：采用自制的腹泻性贝毒软海绵酸（okadaic acid,OA）单克隆抗体与BSA偶联物为胶体金标记底物研究OA胶体金免疫检测技术。制备了各种粒径的胶体金,最终确定粒径30 nm的胶体金作为标记抗OA单克隆抗体效果最佳;制备了半抗原OA与BSA的偶联物,将偶联物包被在硝酸纤维素膜上作为检测带,以羊抗鼠Ig G作为质控带,依据免疫竞争法原理,建立了快速检测OA的免疫层析试纸条方法,该方法在3~5 min即可目测判断结果,灵敏度为6.25 ng/m L;依据工艺流程,发明制备了软海绵酸OA的免疫层析快速检测卡,该卡检测样本中的OA含量阈值为6.25 ng/m L。OA colloidal gold immunoassay technology was studied by adopting self- manufactured diarrhetic shellfish poison,okadaic acid（ OA）,monoclonal antibody and BSA conjugate as the colloidal gold labeled substrate. Colloidal gold was prepared with various particle sizes and the colloidal gold with a particle size of 30 nm was determined as the optimal one for labeling anti-OA monoclonal antibody. The conjugate of hapten OA and BSAwas prepared,immunochromatographic strip method for OA rapid detection was established by coating conjugate on nitrocellulose membrane as inspection band,and taking Goat anti Mouse Ig G as quality control band based on immune competition law. The method can be used in visual judgment in 3-5 min. The sensitivity is 6. 25 ng / m L. According to the process procedure,the immunochromatography rapid assaying card of OA was invented. No instrument and equipment are needed for the card. The OA content threshold value of detecting specimen is 6. 25 ng / m L.

关 键 词：腹泻性贝毒 软海绵酸 单克隆抗体 胶体金 免疫监测 

分 类 号：Q789[生物学—分子生物学]