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Position: Home > Articles > Development and Trait Identification of Rapid Test Blocking ELISA Kit for Sulfadiazine Residues Journal of Henan Agricultural Sciences 2009,38 (11) 122-125+133

磺胺嘧啶残留阻断ELISA试剂盒的研制及鉴定

作  者:
刘庆堂;职爱民;李青梅;柴书军;张磊;邓瑞广;张改平
单  位:
河南省农业科学院河南省动物免疫学重点实验室农业部动物免疫学重点开放实验室
关键词:
磺胺嘧啶;单克隆抗体;阻断ELISA;快速检测试剂盒
摘  要:
基于1株分泌抗磺胺嘧啶(sulfadiazine,SD)高亲和力的SD单克隆抗体(monoclonal antibody,mAb),应用ELISA试验原理研制SD残留快速检测阻断ELISA试剂盒(SD-Kit),并对其特性进行测定。SD-Kit的标准曲线呈典型的S型,相关系数R2=0.9914,符合4参数logit曲线拟合,线性检测范围为0.02~12.92μg/L,灵敏度为0.08μg/L,半数抑制浓度(IC50)为0.54μg/L,检测限为0.08μg/L;鲜奶样和猪尿样的平均添加回收率分别为90.8%、95.8%,平均批内和批间变异系数均低于10%;基质对SD-Kit的检测结果影响不大;SD-Kit与磺胺甲基嘧啶的交叉反应率为2.7%,与其他磺胺类药几乎没有反应性;试剂盒在4℃可保存6个月。
译  名:
Development and Trait Identification of Rapid Test Blocking ELISA Kit for Sulfadiazine Residues
作  者:
LIU Qing-tang,ZHI Ai-min,LI Qing-mei,CHAI Shu-jun,ZHANG Lei,DENG Rui-guang,ZHANG Gai-ping(Henan Key Laboratory for Animal Immunology,Key Laboratory for Animal Immunology of the Ministry of Agriculture,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China)
关键词:
Sulfadiazine;Monoclonal antibody;Blocking ELISA;Rapid test Kit.
摘  要:
A blocking ELISA test kit for detection of sulfadiazine(SD)was developed based on SD monoclonal antibodies(mAb).The calibration curve of the SD-Kit with standard SD inhibitor was typical sigmoid curve fitted to the four parameter logistic equation with the linear detection of 0.02 to 12.92μg/L(R2=0.9914),the sensitivity of 0.08μg/L,the IC50 of 0.54μg/L and the detection limit of 0.08μg/L.The recoveries of SD spiked in milk were 90.8%,in pig urine were 95.8%.The precision and accuracy of the assay as determined by inter-assay and intra-assay coefficient variation was both below 10%.The SD-Kit generally had 2.7% cross-reactivity towards sulfamerazine and little or no cross-reactivity towards other sulfonamides.The dilution solution of SD had no effect on results of SD-Kit.The validity of SD-Kit in 4℃ was above six months.

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