对硫磷酶联免疫吸附分析方法的建立和评价

Development and evaluation of indirect competitive enzyme-linked immunoassay for the parathion

摘要: 通过制备对硫磷的多克隆抗体,建立了对硫磷的间接竞争酶联免疫吸附分析方法(icELISA)。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,获得免疫抗原和包被抗原。使用对硫磷抗血清,经优化条件参数建立了对硫磷的icELISA分析方法。该方法检测线性范围为3.39～533.05ng/mL,最低检测限为0.90ng/mL,对蔬菜样品和水样品的平均添加回收率为75.6%～127.4%,能用于实际样品中对硫磷的检测。

Abstract: Polyclonal antibody (PcAb) against parathion was raised and used to develop an indirect competitive enzyme-linked immunoassay (icELISA) .The active eser method was employed to conjugate the hapten to bovin serum albumin (BSA) and ovalbumin (OVA) as immounogen and coating antigen respectively.Anti-parathion PcAb was used to develope icELISA.The result showed that the linear detection of this icELISA system was 3.39～ 533.05ng/mL, the detection limit was 0.90ng/mL, the average recovery of parathion from vegetables and water samples was 75.6%～127.4%.Therefore, the icELISA established was a pratical method for detection of parathion in real samples.