食品科学

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基于不同半抗原的呋喃唑酮代谢物免疫检测方法的建立与比较

石贤爱1,2,裴世锋1,李向楠1,朱秋享1,叶小军3,樊海平3   

  1. 1.福州大学生物科学与工程学院,福建 福州 350108;2.福建省医疗器械和医药技术重点实验室,福建 福州 350002;
    3.福建省淡水水产研究所,福建 福州 350002
  • 出版日期:2014-04-25 发布日期:2014-05-13
  • 基金资助:

    国家海洋公益性行业科研专项(201205022-3);福建省科技重大专项(2013NZ0003);
    福建省海洋与渔业厅重点项目(闽海渔合同[2010]2-27号)

Establishment and Comparison of Immunoassays for Detecting Furazolidone Metabolite Based on Different Haptens

SHI Xian-ai1,2, PEI Shi-feng1, LI Xiang-nan1, ZHU Qiu-xiang1, YE Xiao-jun3, FAN Hai-ping3   

  1. 1. College of Biological Science and Engineering, Fuzhou University, Fuzhou 350108, China;
    2. Fujian Key Laboratory of Medical Instrument and Pharmaceutical Technology, Fuzhou 350002, China;
    3. Fresh Water Fisheries Research Institute of Fujian Province, Fuzhou 350002, China
  • Online:2014-04-25 Published:2014-05-13

摘要:

为实现呋喃唑酮代谢物3-氨基-2-恶唑烷酮(3-amino-2-oxazolidinone,AOZ)的快速定量检测,制备AOZ- 牛血清蛋白单克隆抗体,建立2 种分别基于抗3-(4-羧基苯亚甲基)-氨基-2-恶唑烷酮(CPAOZ)、抗AOZ的单克 隆抗体酶联免疫检测试剂盒。CPAOZ检测试剂盒在1.0~100.0 ng/mL范围具有较好的线性,IC50值为14.6 ng/mL, 检测限为6.56 ng/mL,回收率为97.6%~101.3%;AOZ检测试剂盒在0.5~12.5 ng/mL范围具有良好线性,IC50值为 3.9 ng/mL,检测限为0.45 ng/mL,回收率为94.1%~97.4%。这2 种检测试剂盒对于其他硝基呋喃类抗生素及其代谢 物均不存在交叉反应。从经济、方便、快速、灵敏等因素来考虑,后者更具有发展前景。

关键词: 3-氨基-2-恶唑烷酮, 半抗原, 单克隆抗体, 酶联免疫检测

Abstract:

Based on the haptens of the furazolidone metabolite 3-amino-2-oxazolidinone (AOZ) and its derivative CPAOZ,
mAb-based ELISA kits for the detection of AOZ were established after the preparation of monoclonal antibodies (mAbs)
against AOZ-BSA. The CPAOZ-based ELISA kit indicated excellent linearity over the range of 1.0–100.0 ng/mL, The IC50
was 14.6 ng/mL, with detection limit of 6.56 ng/mL, and the recovery for CPAOZ was 97.6%–101.3%. The AOZ-based
ELISA kit exhibited excellent linearty over the range of 0.5–12.5 ng/mL, The IC50 was 3.9 ng/mL, with the detection limit
of 0.45 ng/mL, and the recovery for AOZ was 94.1%–97.4%. Neither of the ELISA kits showed cross reactivity with other
nitrofurans or their metabolites. Obviously, the AOZ-based ELISA kit has more promising prospects due to its advantages of
economy, convenience, rapidity and sensitivity.

Key words: 3-amino-2-oxazolidinone (AOZ), hapten, monoclonal antibody (mAb), enzyme-linked immunosorbent assay (ELISA)

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