食品科学 ›› 2017, Vol. 38 ›› Issue (10): 305-309.doi: 10.7506/spkx1002-6630-201710049

• 安全检测 • 上一篇    

磁性化学发光酶免疫法检测猪肉中的氯霉素

王 玲,管 笛,周 欣,高书涛,臧晓欢,魏 颖   

  1. 1.河北农业大学理学院,河北 保定 071001;2.北京市理化分析测试中心,北京 100089
  • 出版日期:2017-05-25 发布日期:2017-05-23
  • 基金资助:
    河北省科技厅科技支撑重点项目(肉类及豆制品中有害物质的残留检测)(14227115D)

Development of Magnetic Bead-Based Chemiluminescent Enzyme Immunoassay for Chloramphenicol Detection in Swine Muscle

WANG Ling , GUAN Di , ZHOU Xin , GAO Shutao , ZANG Xiaohuan , WEI Ying   

  1. 1. College of Science, Hebei Agricultural University, Baoding 071001, China; 2. Beijing Center for Physical and Chemical Analysis, Beijing 100089, China
  • Online:2017-05-25 Published:2017-05-23

摘要: 建立以磁性微球为固定相的化学发光酶免疫法,定量检测猪肉中的氯霉素含量。此方法为在微孔中加入羊 抗鼠磁性微球、样品和抗体,反应一定时间,之后补加酶标抗原,继续反应一定时间后清洗磁珠,加入发光液反应 10 min后分析测定。通过磁珠用量、酶标抗原量、抗体量和反应模式的优化,成功建立了一种高灵敏度、高准确度 的检测猪肉中氯霉素的磁性化学发光酶免疫方法。该方法的检出限为0.013 μg/kg,线性范围在0.060~2.421 μg/kg之 间,回收率在85.24%~93.57%之间,相对标准偏差小于11.5%。

关键词: 化学发光酶免疫法, 氯霉素, 磁性微球, 猪肉

Abstract: A chemiluminescent enzyme immunoassay based on magnetic beads was developed for quantitative determination of chloramphenicol (CAP) residues in pork samples. Sample, anti-chloramphenicol antibody, and magnetic beads coated with goat anti-mouse antibody were mixed on the microwell plate. After reacting for a certain period of time, CAP-horseradish peroxidase (HRP) was added and incubated. Then chemiluminescence substrate solution was added after washing the magnetic beads three times. Ten minutes later, the results were recorded by a microplate reader. Besides, the amount of magnetic beads coated with goat anti-mouse antibody, CAP-HRP, antibody concentration and reaction pattern were optimized. A sensitive and accurate magnetic bead-based chemiluminescent enzyme immunoassay for CAP was successfully developed. The limit of detection was 0.013 μg/kg, and good linear relationship was observed in the concentration range between 0.060 and 2.421 μg/kg. The recoveries of spiked pork samples varied from 85.24% to 93.57%, with relative standard deviation of less than 11.5%.

Key words: chemiluminescent enzyme immunoassay, chloramphenicol, magnetic beads, swine muscle

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