摘要
为了建立比较间接竞争化学发光酶联免疫分析法（ic-CLEIA）与直接竞争化学发光酶联免疫分析法（dc-CLEIA）检测家禽饲料中玉米赤霉烯酮（zearalenone,ZEN）的方法,采用二因子交叉试验优化确定最佳包被原浓度和抗体稀释度,建立ic-CLEIA和dc-CLEIA。结果显示：icCLEIA在0.05-5 ng/m L范围内线性关系良好,线性回归方程为y=-0.383 5x＋0.417 9（R^2=0.994 6）,灵敏度（IC50）为0.611 ng/m L,检出限（LOD）为15 pg/m L,平均批内和批间变异系数为1.79%和1.02%,家禽饲料添加样品回收率为91.9%-112.1%,变异系数小于10.5%;dcCLEIA在0.025-5 ng/m L范围内线性关系良好,线性回归方程为y=-0.386 5x＋0.334 1（R^2=0.991 8）,灵敏度（IC50）为0.372 ng/m L,检出限（LOD）为3 pg/m L,平均批内和批间变异系数为1.16%和1.20%,添加样品回收率为87.3%-119.3%,变异系数小于9.8%。结果表明：两种方法精确,灵敏度高,均可作为定量检测ZEN的有效手段;与ic-CLEIA相比,dc-CLEIA更适用于检测家禽饲料中的ZEN。
Indirect competitive chemiluminescence enzyme immunoassay（ic-CLEIA） and direct competitive chemiluminescence enzyme immunoassay（dc-CLEIA） were established here for detection of zearalenone in poultry feed. The two assays were then compared. The optimum concentration of ZEN-BSA and dilution of monoclonal antibody（or Mc Ab-HRP） were optimized by a two factor cross test to establish ic-CLEIA and dc-CLEIA. Under the optimized condition,the developed ic-CLEIA had good linearity within the range of 0. 05 - 5. 0 ng/m L,and the calculated regression equations were y =-0. 383 5 x＋ 0. 417 9（R^2= 0. 994 6）. The developed dc-CLEIA had good linearity within the range of 0. 025 - 5. 0 ng/m L,and the calculated regression equations were y =-0. 386 5 x＋0. 334 1（R^2= 0. 991 8）. From the regression equations,the developed ic-CLEIA was estimated as 50% inhibitory concentration（IC50） value（0. 611 ng/m L） with average coefficients of variation of intra-and inter-assay being1. 79% and 1. 02% respectively,and a limit of detection of 15 pg/m L. The developed dc-CLEIA was estimated as 50% inhibitory concentration（IC50） value（0. 372 ng/m L） with average coefficients of variation of intra-and inter-assays being 1. 16% and 1. 20% respectively,and a limit of detection of 3 pg/m L. The recovery rates of the two assays were 91. 8% - 112. 1%and 87. 3% - 119. 3% respectively for fortified samples with average coefficients of variation being 10. 5% and 9. 8%respectively. Both the methods were precise and sensitive,and could be used as an effective means for quantitative detection of ZEN. Compared with ic-CLEIA,dc-CLEIA was more suitable for the follow-up detecting study of ZEN in poultry feed.
出处
《畜牧与兽医》
北大核心
2018年第2期122-130,共9页
Animal Husbandry & Veterinary Medicine
基金
国家重点研发计划(2016YFD0500800-10)
上海市技术标准专项(13DZ0502701)
关键词
玉米赤霉烯酮
间接竞争化学发光酶联分析法
直接竞争化学发光酶联分析法
家禽饲料
zearelanone （ZEN）
indirect competitive chemiluminescence enzyme immunoassay （ic-CLEIA）
direct competitive chemiluminescence enzyme immunoassay （dc-CLEIA）
poultry feed
