赭曲霉毒素A免疫学检测方法的研究

江涛; 李凤琴; 王环宇; 田静; 高秀芬; 宫慧之; 王玉平; 计融

doi:10.11847/zgggws2004-20-05-26

赭曲霉毒素A免疫学检测方法的研究

Study on immunoassay for ochratoxin A

摘要

摘要:
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2~500ng/ml, 线性方程Y=0.272X+1.07(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60.71%, 最大值为8.26μg/kg; 大米样品的污染率为17.86%, 最大值为3.44μg/kg。
结论该方法简单、快速、灵敏, 完全可以满足实际工作的需要。

Abstract:
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 8.26μg/kg; the contamination rate of rice as 17.86%, the maximum level was 3.44μg/kg.
Conclusion The detection method was simple, fast sensitive and can meet the demand of practical work.

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