华北农学报 ›› 2009, Vol. 24 ›› Issue (2): 214-217. doi: 10.7668/hbnxb.2009.02.044

• 论文 • 上一篇    下一篇

磺胺甲噁唑残留检测阻断ELISA试剂盒的研制

刘宣兵1,2, 张改平1, 王斌3, 滕曼1, 邓瑞广1, 侯玉泽2, 邢广旭1, 杨继飞1   

  1. 1. 河南省农业科学院动物免疫学重点实验室, 河南郑州 450002;
    2. 河南科技大学 食品与生物工程学院, 河南洛阳 471003;
    3. 山东诸城城市供排水管理处, 山东诸城 262200
  • 收稿日期:2008-12-11 出版日期:2009-04-28
  • 通讯作者: 张改平( 1960- ), 男, 河南内黄人, 研究员, 博士生导师, 主要从事动物免疫学和兽药残留研究。
  • 作者简介:刘宣兵( 1982- ), 男, 山东诸城人, 硕士, 主要从事兽药残留检测与食品安全研究.
  • 基金资助:
    "十一五"国家科技支撑计划(2006BAK02A21)

Development of Rapid Test Blocking ELISA Kit for Sulfamethoxazole Residues

LIU Xuan-bing1,2, ZHANG Gai-ping1, WANGBin3, TENGMan1, DENG Rui-guang1, HOU Yu-ze2, XING Guang-xu1, YANGJi-fei1   

  1. 1. Henan Key Laboratory for Animal Immunology, Agriculture Academy of Henan Province, Zhengzhou 450002,China;
    2. Food and Bioengineering College, Henan University of Science and Technology, Luoyang 471003, China;
    3. Zhucheng City Admintsirative Office of Supply-discharge Water, Zhucheng 262200, China
  • Received:2008-12-11 Published:2009-04-28

摘要: 在建立磺胺甲噁唑单克隆抗体(SMZmAb)杂交瘤细胞株和阻断ELISA方法的基础上,研制出SMZ残留快速检测试剂盒(SMZ-Kit),并对其性能进行了测定.结果表明,SMZ-Kit的标准曲线呈典型的S型,符合参数logit曲线拟合,相关系数R2=0.990 1,检测范围为1.0~128.0 μg/L,灵敏度为0.73μg/L,半数抑制浓度(IC50)为11.5 μg/L,检测限为1.0μg/L;饲料样、牛奶样的平均添加回收率为83.9%,83.3%,平均批内和批间变异系数均<15%;SMZ-Kit与磺胺嘧啶的交叉反应(CR%)为2.88%,与其他磺胺类药物无交叉反应;试剂盒在℃可保存6个月.

关键词: 磺胺甲噁唑, 单克隆抗体, 阻断EUSA, 快速检测试剂盒

Abstract: A ELISA kit for detection of Sulfamethoxazole (SMZ-Kit)was developed with generating hybridoma lines that secrete SMZ monoclonal antibodies (SMZmAb) and the method of blocking ELISA,its traits were identified. The calibration curve of SMZ-Kit was typical sigmoid curve fitted to the four parameter logistic equation with the linear detection of 1.0 to 128.0μg/ L (R2 = 0.9901),the sensitivity of 0.73μg/ L,the IC50 of 11.5μg/ L and the detection limit of 1.0μg/L.The recoveries of SMZ spiked in feed were 83.9 % and in milk were 83.3 %.The precision and accuracy of the assay as determined by inter-assay and intra-assay coefficient variation were below 15 %.The SMZ - Kit had 2.88 % cross-reactivity towards Sulfadiazine (SD) and little or no cross-reactivity towards other Sulfonamides.The validity of SMZ-Kit in 4℃was above six months.

Key words: Sulfamethoxazole, Monoclonal antibody, Blocking ELISA, Rapid test kit

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引用本文

刘宣兵, 张改平, 王斌, 滕曼, 邓瑞广, 侯玉泽, 邢广旭, 杨继飞. 磺胺甲噁唑残留检测阻断ELISA试剂盒的研制[J]. 华北农学报, 2009, 24(2): 214-217. doi: 10.7668/hbnxb.2009.02.044.

LIU Xuan-bing, ZHANG Gai-ping, WANGBin, TENGMan, DENG Rui-guang, HOU Yu-ze, XING Guang-xu, YANGJi-fei. Development of Rapid Test Blocking ELISA Kit for Sulfamethoxazole Residues[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(2): 214-217. doi: 10.7668/hbnxb.2009.02.044.

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