化学发光-酶联免疫高灵敏检测洛美沙星

J4 ›› 2011, Vol. 46 ›› Issue (11): 12-16.

化学发光-酶联免疫高灵敏检测洛美沙星

曹照真¹,卢圣欣¹,郗日沫^2*

1.山东大学化学与化工学院, 山东济南 250100; 2.南开大学药学院, 天津 300071

收稿日期:2011-05-23 出版日期:2011-11-20 发布日期:2011-11-30
通讯作者: 郗日沫(1962- ),男,博士,教授,研究方向为无机及分析化学. Email: xirimo2000@yahoo.com
作者简介:曹照真(1968- ),女,硕士,工程师, 研究方向为无机及分析化学. Email: zzcao2002@sdu.edu.cn
基金资助:
中国国家自然科学基金资助项目(20675048); 国家高技术研究计划发展(863计划)资助项目(2007AA10Z435, 2007AA06A407); 山东省自然科学基金资助项目(ZR2009BM004)

A highly sensitive chemiluminescent enzymelinked immunosorbent assay for the determination of lomefloxacin

CAO Zhao-zhen¹, LU Sheng-xin¹, XI Ri-mo^2*

1. School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, Shandong, China;
2. College of Pharmacy, Nankai University, Tianjin 300071, China

Received:2011-05-23 Online:2011-11-20 Published:2011-11-30

摘要/Abstract

摘要：

建立了洛美沙星的化学发光-酶联免疫分析方法。实验对包被抗原的度盘浓度、洛美沙星抗体及辣根过氧化物酶标记的羊抗兔二抗的稀释倍数、酶标板的种类等进行了优化。在最佳实验条件下,测得的IC₅₀值为0.22ng／mL, 检测限为0.032ng／mL。该方法具有良好的特异性,在检测的10种代表性的喹诺酮类药物中,仅诺氟沙星和氟罗沙星交叉率高于1％(分别为:6.88％和2.16％)。将该方法应用于脱脂牛奶中添加洛美沙星回收率的测定,批内和批间回收率分别为:88.7％～103.4％和97.8％～107.2％,相应变异系数分别为:6.3％～8.6％和2.4％～8.0％。

关键词: 抗体;化学发光;酶联免疫;药物残留;洛美沙星

Abstract:

A chemiluminescent enzyme-linked immumosorbent assay (CL-ELISA) based on polyclonal antibody for lomefloxacin was developed. Assay conditions, including the concentration of coating antigen, the dilution degree of the primary antibody and IgG-HRP, and the type of micro-titer plate. were studied. The optimized CL-ELISA presented high sensitivity with an IC₅₀value of 0.22ng／mL and detection limit of 0.032ng／mL. Good specificity was acquired with only norfloxacin and fleroxacin showing a percentage of cross-reactivity higher than 1％ (6.88％ and 2.16％ respectively) among the ten detected representative quinolones. The method developed was applied in the detection of lomefloxacin recovery in spiked milk. The average recovery rates were in the ranges of 88.7％～103.4％ and 97.8％～107.2％ for intra- and inter-assay respectively while the corresponding coefficient of variation (CV％) were in the ranges of 6.3％～ 8.6％ and 2.4％～ 8.0％.

Key words: antibody; chemiluminesence; ELISA; drug residue; lomefloxacin

曹照真1,卢圣欣1,郗日沫2*. 化学发光-酶联免疫高灵敏检测洛美沙星[J]. J4, 2011, 46(11): 12-16.

CAO Zhao-zhen1, LU Sheng-xin1, XI Ri-mo2*. A highly sensitive chemiluminescent enzymelinked immunosorbent assay for the determination of lomefloxacin[J]. J4, 2011, 46(11): 12-16.

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