饲料中喹乙醇ELISA检测试剂盒的研制及性能分析  

Development and Performance of ELISA Kit for Rapid Detection of Olaquindox in Feed

作　　者：张景艳[1] 王磊[1] 李建喜[1] 张宏 张凯[1] 杨志强[1] 王学智[1] 秦哲[1] 孟嘉仁[1] 

机构地区：[1]中国农业科学院兰州畜牧与兽药研究所农业部新兽药工程重点实验室甘肃省中兽药工程技术研究中心,兰州730050 [2]甘肃省动物卫生监督所,兰州730000

出　　处：《畜牧兽医学报》2013年第8期1297-1304,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：中央级公益性科研院所基本科研业务费专项资金项目(BRF090303);甘肃省科技支撑计划项目(1011NKCA048)

摘　　要：本研究旨在研制快速检测饲料中喹乙醇的ELISA试剂盒(OLA-Kit)。作者用喹乙醇单克隆抗体(OLAmAb)建立间接竞争ELISA方法,组装OLA-Kit,并对试剂盒的性能进行评估。结果表明:OLA-Kit的标准曲线呈典型的反S型,相关系数R2=0.976 7,线性范围为1～243μg·L-1,半数抑制浓度(IC50)为(14.29±3.75)μg·L-1,检测限为0.79μg·L-1,定量限为4.0μg·L-1,临界值(猪饲料)为83.37μg·g-1,基质对该试剂盒的检测结果影响不大;猪、鸡、鱼饲料的不同浓度添加回收率均大于74.40%,平均批内和批间变异系数均小于15%;试剂盒与喹胺醇的交叉反应率(CR%)为17.93%,与MQCA、QCA及其他喹喔啉类药物几乎无反应性,可在2～8℃保存6个月以上;OLA-Kit与HPLC法测定猪饲料中喹乙醇添加回收率没有显著差异(P≥0.05)。本研究成功研制出敏感、特异、准确的喹乙醇ELISA检测试剂盒,可用于饲料中该药物的快速检测和批量筛查。The aim of this study was to develop a rapid detection kit for olaquindox（OLA-Kit） in animal feed.The OLA monoclonal antibody（OLA mAb） was applied to assemble the indirect competitive ELISA kit and its characterization was determined.The results showed that the calibration curve of OLA-Kit was a typical sigmoid curve with the linear detection of 1-243 μg· L-1,and R2 was 0.976 7.The kit has a half inhibition concentration（IC50） of（14.29±3.75） μg·L-1,the detection limit of 0.79 μg·L-1,the quantitative limit of 4.0 μg·L-1,the cut-off value of 83.37 μg·g-1 for pannage,and the matrix has no effect on the results of the OLA-Kit;The recoveries of OLA in pig feed,chicken feed and fish feed were all above 74.40%,and the inter-assay and intra-assay coefficient variation were all below 15%;The OLA-Kit had 17.93% cross-reactivity towards Kuianchun and little or no cross-reactivity towards MQCA,QCA and other quinoxaline drugs.The validity of kit in 2-8 ℃ was above six months.There was no significant difference of the recovery of OLA in pig feed between OLA-Kit and HPLC（P≥0.05）.In conclusion,the OLA-Kit was sensitive,specific and accurate,could be used for the rapid determination and extensive screening of OLA in animal feed.

关 键 词：喹乙醇 单克隆抗体 竞争ELISA 试剂盒 饲料 

分 类 号：S859.796[农业科学—临床兽医学]