抗腹泻性贝毒软海绵酸胶体金免疫快速检测技术研究

摘要

OA colloidal gold immunoassay technology was studied by adopting self-manufactured diarrhet-ic 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 IgG 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/mL.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/mL.%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IgG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/mL;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/mL.