磺胺间甲氧嘧啶残留阻断ELISA试剂盒的研制及其初步应用  

Develepment and trait identification of rapid test blocking ELISA Kit for sulfamonomethoxine residues

作　　者：职爱民[1] 李青梅[1] 刘庆堂[1] 邢广旭[1] 杨继飞[1] 柴书军[1] 邓瑞广[1] 张改平[1] 

机构地区：[1]河南省农业科学院农业部动物免疫学重点开放实验室,河南郑州450002

出　　处：《西北农林科技大学学报（自然科学版）》2009年第10期37-42,共6页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家"十一五"科技支撑计划项目(2006BAK02A21)

摘　　要：【目的】研制磺胺间甲氧嘧啶（Sulfamonomethoxine,SMM）残留快速检测阻断ELISA试剂盒（SMM-Kit）,为有效治理磺胺间甲氧嘧啶的滥用,保障动物源性食品安全提供技术支撑。【方法】在成功研制SMM单克隆抗体（Monoclonal antibody,mAb）的基础上,应用阻断ELISA试验原理研制SMM-Kit,并对其特性进行了系统测定。【结果】SMM-Kit标准曲线呈典型的S型,相关系数R^2=0.992 8,符合4参数logit曲线拟合,线性检测范围为1.33～389.9μg/L,灵敏度为3.57μg/L,半数抑制浓度（IC50）为17.07μg/L。牛奶样和猪尿样的平均添加回收率分别为91.5%和92.0%,平均批内和批间变异系数均低于15%;基质对SMM-Kit检测结果的影响不大;SMM-Kit与其他磺胺类药物的交叉反应性〈1.7%,表明其与其他药物几乎无交叉反应性;试剂盒在4℃可保存6个月。【结论】成功地研制出了SMM-Kit,在牛奶和猪尿液样品中的初步应用证明,该试剂盒灵敏度、准确度高,特异性强,可低温保存6个月。[Objective] A blocking ELISA test Kit for detection of Sulfamonomethoxine（SMM）was developed to offer technological support for effective governance＇of the abuse of sulfamonomethoxine and protection of animal derived food safety. [Method] Based on a high sensitivity monoclonal antibody of SMM,a rapid test ELISA Kit was established and identified by blocking EIASA. [Result] The calibration curve of the SMM-Kit with standard SMM inhibitor was typical sigmoid curve fitted to the four parameter logistic equation with the linear detection of 1. 33-389. 9 μg/L（R^2 = 0. 992 8）, the sensitivity of 3.57 μg/L,the IC50 of 17. 07 μg/L. The recoveries of SMM spiked in milk were 91.5%, in pig urine 92.0%. The precision and accuracy of the assay as determined by inter-assay and intra-assay coefficient variation was both below 15%. The SMM-Kit generally had 1.7% cross-reactivity towards sulfamerazine and had little or no cross-reactivity towards other compounds. The dilution solution of SMM had no effect on results of SMM-Kit. The validity of SMM-Kit in 4 ~C was above six months. [Conclusion] A blocking SMM-Kit was established and identified. The successful dues of milk and Urine showed that the SMM-Kit had a preliminary application in detection for SMM resihigh sensitivity, accuracy, specificity and the validity at low temperature was above six months.

关 键 词：磺胺间甲氧嘧啶 单克隆抗体 阻断ELISA 快速检测试剂盒 

分 类 号：S859.84[农业科学—临床兽医学]