化学发光免疫法检测食品中恩诺沙星残留的研究

龚云飞; 邹晓楠; 张露露; 吴莹莹; 戴明雁; 陈宗伦; 张明洲

doi:10.13386/j.issn1002-0306.2014.04.046

化学发光免疫法检测食品中恩诺沙星残留的研究

中国计量学院生命科学学院浙江省生物计量及检验检疫技术重点实验室
杭州迪恩科技有限公司

基金项目:

浙江省重点科技创新团队(2010R50028)；浙江省科技成果转化项目(2011E61018)；浙江省国境安全检验检疫科技创新服务平台(2006C17014)；浙江省重大科技专项重点项目(2012C12013-3)；

详细信息

作者简介:
龚云飞 (1984-) , 硕士研究生, 主要从事食品安全快速检测技术方面的研究。;

中图分类号: TS207.3
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出版历程
- 收稿日期: 2013-08-01

Study on detecting enrofloxacin residues in food by chemiluminescence immunoassay

College of Life Science, China Jiliang University, Zhejiang Provincial Key Laboratory of Bio-metrology, Inspection and Quarantine
Hangzhou DNA Sci-tech Co., Ltd.

摘要

摘要: 建立了基于多克隆抗体的化学发光免疫检测技术,用于测定动物源性食品中恩诺沙星残留含量。反应条件优化结果为:抗体包被最佳稀释倍数为64000倍,最佳酶标抗原稀释倍数为256000倍。结果显示:该方法的回归曲线方程为y=-13.898x+110.38（R2=0.9893）,检测线性范围为3⁸10pg/mL,IC50为69.63pg/mL,IC10为1.41pg/mL;鸡肉、鱼肉、虾肉、蜂蜜、牛奶空白组织样品中恩诺沙星的最低检测限分别为3.76、4.59、2.85、2.65、4.20pg/mL,最低定量限分别为6.13、7.35、3.57、3.73和6.48pg/mL,回收率在88.28%¹02.6%,板内和板间的平均变异系数分别为2.61%和4.71%。表明本研究建立的化学发光免疫检测方法满足动物源性食品恩诺沙星残留的检测要求。
- 恩诺沙星 /
- 化学发光免疫检测技术 /
- 食品安全
Abstract: It was developed a chemiluminescence immunoassay (CLIA) method based on polyclonal antibody to detect the enrofloxacin residue in animal food. The CLEIA conditions were optimized. The optimal concentration of coating antibody dilution was 1∶64000 and the enrofloxacin markered enzymes dilution was 1∶256000. The results showed that:with an concentration range from 3 to 810pg/mL, the regression equation was y=-13.898x+ 110.38 (R2=0.9893) . The 50% inhibition percentage (IC50) and sensitivity (IC10) was 69.63pg/mL and 1.41pg/mL, respectively. Actual sample testing experiment results showed:in chicken, fish, shrimp, honey and milk samples, the detection limits was 3.76, 4.59, 2.85, 2.65 and 4.20pg/mL, respectively. The limit of quantity was 6.13, 7.35, 3.57, 3.73 and 6.48pg/mL, respectively. Recoveries were between 88.28% ¹02.6% when ENR was spiked to samples at 50¹000pg/g (pg/mL) . The mean intra-assay variability and inter-assay was 2.61% and 4.71%, respectively. The CLIA which we stablished could be used to detect enrofloxacin residues in animal food.
- enrofloxacin /
- chemiluminescence immunoassay /
- food safety

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参考文献(19)

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