赭曲霉毒素A免疫学检测方法的研究

Study on immunoassay for ochratoxin A

目的 建立敏感、特异和快速针对赭曲霉毒素A的酶联免疫吸附试验检测方法 ,研制具有我国自主知识产权的快速检测试剂盒。方法 利用B细胞杂交瘤技术 ,建立能够分泌抗赭曲霉毒素A单克隆抗体的杂交瘤细胞株 ,并获得抗赭曲霉毒素A单克隆抗体。结果 建立赭曲霉毒素A的间接竞争抑制性酶联免疫吸附试验测定方法(ELISA)。该方法的最低检出浓度为 0 5ng/ml,线性范围 2～ 5 0 0ng/ml,线性方程Y =0 2 72X +1 0 7(r =0 9978)。方法的加标回收率为 79 0 %～ 119 7%。利用该方法对北京市售的大米、小麦样品进行检测。结果表明 ,小麦样品的污染率为 6 0 71% ,最大值为 8 2 6 μg/kg ;大米样品的污染率为 17 86 % ,最大值为 3 4 4 μg/kg。结论 该方法简单、快速、灵敏 。

Objective To develop a sensitive,specific and rapid enzyme-linked immunosorbent assay for detection of ochratoxin A(OA).To develop rapid detection kits that possessing patent of our country.Methods To produced hybridoma cell lines excreting monoclonal antibodies against OA by using B cell hybridom technique.And to obtain the monclonal antibodies against OA.Results The indirect competitive inhibition enzyme-linked immunosorbant assay(ELISA) was developed for detection OA in rice and wheat.The limit of detection concentration of OA was 0.5?ng/ml.The linear range of standard curve was between 2-500?ng/ml,the linear equation was Y=-0.272X+1.07( r =0 997?8).The recovery of OA was between 79 0%-119.7%.The OA contamination level of rice and wheat that were sold in Beijing was surveyed by using the method.Contamination rate of wheat was 60.71%,the maximum level was 8.26?μg/kg;the contamination rate of rice as 17.86%,the maximum level was 3.44?μg/kg.Conclusion The detection method was simple,fast sensitive and can meet the demand of practical work.