对硫磷化学发光酶联免疫吸附分析方法的建立和评价  

Development of an Indirect Competitive Chemiluminescence Enzyme-linked Immunoassay for Parathion

作　　者：邓浩[1] 孔德彬 杨金易[1] 徐振林[1] 沈玉栋[1] 杨星星[1] 孙远明[1] 

机构地区：[1]广东省食品质量安全重点实验室,华南农业大学食品学院,广州510642 [2]国家肉制品质量监督检验中心(漯河),漯河462000

出　　处：《分析化学》2013年第2期247-252,共6页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(No.30901005);国家科技支撑计划(No.2012BAD31B03);广东省科技计划(Nos.2011A090200029,2010A032000001-4,2010A090200084)资助项目

摘　　要：建立了基于多克隆抗体的对硫磷间接竞争化学发光酶联免疫吸附分析方法(icCLEIA)。以三氯硫磷为原料,经三步取代反应合成对硫磷半抗原,通过活泼酯法将半抗原分别与牛血清蛋白(BSA)和卵清蛋白(OVA)偶联,制备免疫抗原和包被抗原。经免疫新西兰大白兔,获得对硫磷抗血清。通过优化条件参数,建立了对硫磷的icCLEIA分析方法。本方法的检测线性范围为0.24～15.83!g/L;半抑制浓度IC50为1.14!g/L;检出限为0.09!g/L;对蔬菜样品和水样品的平均添加回收率为74.6%～121.0%。本方法可用于实际样品中痕量对硫磷残留检测。Polyclonal antibody （PcAb） against parathion was raised and used to develop an indirect competi- tive chemiluminescence enzyme-linked immunoassay （ieCLEIA）. The hapten was prepared from thiophosphor- yl chloride after a three-step substitution reaction. The hapten was coupled to bovine serum albumin （BSA） and ovalbumin（OVA） as immounogen and coating antigen respectively by active ester method. New Zealand rabbits were immunized by the immunogen to obtain anti-parathion polyclonal antibody. Several parameters that might affect ieCLEIA performance were carefully optimized. Under the optimum conditions, the linear range of the developed ieCLEIA was 0.24-15.83 μg/L, the IC50 was 1.14 9g/L and the limit of detection was 0.09μg/L. The average recovery of parathion from spiked vegetables and water samples ranged from 74.6% to 121.0%. In conclusion, the icCLEIA is a practical method for trace detection of parathion in real samples.

关 键 词：对硫磷 化学发光酶联免疫吸附分析 农药残留 

分 类 号：O657.3[理学—分析化学]