赭曲霉毒素A直接竞争ELISA试剂盒的研制

在多克隆抗体的基础上研制了赭曲霉毒素A(OTA)直接竞争酶联免疫检测(cd-ELISA)试剂盒.在0 ～ 10 ng/mL范围内,该试剂盒50％抑制率(IC50)为1.09 ng/mL,检测灵敏度(IC15)为0.08 ng/mL;与赭曲霉毒素B、C的交叉反应率分别为6.28％和0.16％,而与黄曲霉毒素B1等生物毒素未见有交叉反应；板内与板间平均变异系数分别为2.21％和2.79％；花生、玉米和玉米粉3种样品中OTA的检测低限分别为1.71,1.26和1.85 μg/kg,平均添加回收率在83.80％～ 91.40％;与HPLC检测方法具有较高的相关性,相关系数(R2)分别为0.94、0.88和0.90;检测时间只需20 min,可用于花生、玉米及玉米粉中OTA的批量快速筛查.

Development of a Direct Competitive ELISA Kit for Rapid Detection of Ochratoxin A

A direct competitive enzyme-linked immunosorbent assay(cd-ELISA) kit based on ochratoxin A(OTA) polyclonal antibodies was developed.In the 0 ～10 ng/mL range,the OTA ELISA kit had a good sensitivity with an IC50 of 1.09 ng/mL and IC15 of 0.08 ng/mL,showed 6.28%,0.16% cross-reactivitives to ochratoxin B and C respectively,and scarcely showed cross-reactivity to others.The mean variation coefficient of intra-assay and inner-assay were 2.21% and 2.79%.The limit of detection in peanut,maize and maize flour samples were 1.71,1.26 and 1.85 μg/kg,respectively,and spiked recovery ranged from 83.80% to 91.40%.Compared with HPLC,the correlation coefficients(R2) were 0.94、0.88 and 0.90 for peanut,maize and maize flour,respectively.And the detection time of the kit is 20 minutes.It is indicated that the OTA ELISA kit in this paper was applicable to rapid preliminary screening of OTA.