摘要
本研究依据直接竞争ELISA原理建立了齐帕特罗一步法ELISA试剂盒。以齐帕特罗与载体蛋白偶联物作为免疫原免疫新西兰大白兔制得齐帕特罗多克隆抗体,棋盘包被法确定其最佳抗体包被浓度、酶标抗原工作浓度、包被条件、反应时间、底物显色时间等,并对试剂盒的各项技术指标进行确认。结果表明,成功组装了齐帕特罗一步法ELISA试剂盒,并建立了尿液、饲料、奶粉和奶汁,以及组织等样品的前处理方法,检测限均远低于1μg/kg。该试剂盒线性检测范围为0.15～10ng/mL,IC50浮动范围0.43～0.79ng/mL,样品板内、批内、批间的变异系数均小于15%,平均回收率在70%～110%之间,与其同类药物的交叉反应率均小于0.1%。提示,本试验研制的试剂盒重复性、特异性、稳定性等各项指标均符合技术要求,可用于动物源性食品中齐帕特罗药物残留的检测。
This study established a competitive enzyme-linked immunosorbent assay（ELISA） method for detecting zilpaterol in animal derived food.Zilpaterol polyclonal antibody was obtained by immunizing New Zealand rabbit with zilpaterol-KLH conjugates.Checkerboard titration method was used to determine the optimum concentration of antibody and enzyme-labelled antigen.Coating condition,reaction time and the color developing time of substrate were also detected.The results showed that an ELISA kit with a linear sensitivity ranged from 0.15~10 ng/mL was successfully established.Half inhibitory concentration（IC50） ranged between 0.43~0.79 ng/mL.The recovery rates for spiked zilpaterol in urine,feed,milk powder,milk and tissue were 70%~110% with less than 15% coefficient variations（CV）.The cross-reaction for other similar medicines was less than 0.1%.The ELISA kit established in current study for detecting zilpaterol in animal derived food and showed the good reproducibility,stability and specificity,hence it could be used to detect the zilpaterol residues in animal derived food.
出处
《中国畜牧兽医》
CAS
北大核心
2013年第3期242-247,共6页
China Animal Husbandry & Veterinary Medicine
基金
生猪及健康肉安全快速检测关键技术研究与应用(11120206A-1)
关键词
动物源性食品
ELISA
齐帕特罗
animal derived food
enzyme-linked immunosorbent assay
zilpaterol
