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动物组织中盐酸克伦特罗残留ELISA检测试剂盒的研制

Development of an enzyme linked immunosorbent assay kit for clenbuterol residues in animal tissues

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【作者】 李平潘荣生周洪斌肖震方原民宓晓黎李利东丁贵平袁建兴成恒嵩

【Author】 LI Ping,PAN Rong-sheng,ZHOU Hong-bing,XIAO Zhen,FANG Yuan-min,MI Xiao-li,LI Li-dong,DING Gui-ping,YUAN Jian-xing,CHENG Heng-song(Zhenjiang Entry-exit Inspection and Quarantine Bureau,Zhenjiang 212008,China)

【机构】 镇江出入境检验检疫局江苏省微生物研究所江苏省微生物研究所 江苏镇江212008江苏镇江212008无锡214063

【摘要】 目的本研究采用直接竞争酶联免疫吸附试验(ELISA)对动物组织中盐酸克伦特罗(CBL)残留量进行快速筛选检测。方法将CBL与蛋白质载体偶联制备了CBL-BSA和HRP-CBL结合物,以CBL-BSA免疫实验兔获得特异性抗体,并成功建立了CBL残留直接竞争ELISA检测方法及检测试剂盒。结果检测线性范围0.1-62.5 ng/mL,检测低限为0.10 ng/mL;尿、肝、肉和饲料的加标回收率为60.3%-115.0%;与英国Randox试剂盒对比测试结果无显著差异;重现试验变异系数为3.4%-7.2%;试剂盒贮藏在2-8℃5个月和30℃7 d质量稳定。结论本试剂盒具有操作简便、快速、灵敏等特点。

【Abstract】 Objective To rapid screen clenbuterol(CBL) residues in animal tissues by direct competitive ELISA.Methods BSA-CBL and HRP-CBL conjugates were prepared by using CBL coupled to bovine serum albumin and horseradish peroxidase,respectively.Specific antibodies against CBL were prepared by immunizing rabbits with CBL-BSA.ELISA kit has been developed for detecting CBL residues.Re-sults The linear detection range was from 0.1 ng/mL to 62.5 ng/mL with the lowest detection limit of 0.10 ng/mL.The recovery of CBL in spiked urine,liver,meat and feedstuff was ranged from 60.3% to 115.0%,which did not show any significant difference with the detection results of England Randox kit.Coefficient of variation for repeatability tests of standard curve and spiked samples was between 3.4% and 7.2%.The quality of kit stored at 2 ℃-8 ℃ for five month and 30 ℃ for seven day was reliable.Conclusion This testing kit is simple,rapid and sensitive.

【基金】 国家质量监督检验检疫总局科研成果(G027-2004)
  • 【文献出处】 免疫学杂志 ,Immunological Journal , 编辑部邮箱 ,2006年S1期
  • 【分类号】S859.84
  • 【被引频次】12
  • 【下载频次】314
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