摘要
目的:建立检测食品中赭曲霉毒素(OA)快速灵敏化学发光酶免疫方法。方法:采用棋盘滴定法确定OA-BSA抗原的包被质量浓度、包被量以及一抗和二抗的工作稀释度,建立间接竞争抑制曲线,确定线性范围、检出限和回收率。结果:确定的检测工作条件为:OA-BSA最佳包被质量浓度60ng/mL,抗OA单克隆抗体的最佳工作稀释度1:400,校正曲线线性范围6~400ng/mL,赭曲霉毒素的检出限为0.02ng/mL,50%抑制质量浓度145ng/mL,在100~2000ng/g添加水平的回收率范围为83.6%~105.8%。用所建方法对质控样品进行了分析,检测结果符合率达到100%。结论:所建方法准确、灵敏,适用于食品中OA的筛选。
Objective: To establish a sensitive, specific and rapid chemiluminescent immunoassay (CLIA) method for the detection of ochratoxin A (OA) in food. Methods: The optimal conditions of CLIA for OA detection including the concentration of OA-bovine serum albumin (BSA) conjugate, the volume of OA-BSA solution for immobilization, the working concentrations of both antibodies against OA and a horseradish peroxidase-labeled anti-antibody, were explored by chessboard titration experiments. The linear range and the limit of detection (LOD) were studied as well. Certified reference material (CRM) of corn samples containing OA obtained from Food Analysis Performance Assessment Scheme (FAPAS) were analyzed by the developed CLIA method. Results: The optimal CLIA conditions for OA detection were 0.06μg/mL coating concentration of OA-BSA, 100μL/well of coating volume and 1:400 of working titer of the antibody against OA. The linear range of the developed CLIA was 6-400 ng/mL. The half inhibitory concentration was 145 ng/mL and the LOD was 0.04μg/kg for OA in food samples. The recovery rate was in the range of 66%-124% with the spiked sample level of 0.1-2μg/kg. CRM corn samples were analyzed and the results were consistent with the real values. Conclusion: The developed CLIA method was simple, fast, sensitive and can be employed in the screening of OA in foods.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2010年第24期432-435,共4页
Food Science
基金
国家"863"计划项目(2007AA10Z423)
关键词
赭曲霉毒素
化学发光酶免疫分析
检测
ochratoxin A(OA)
chemiluminescent immunoassay (CLIA)
detection