化学发光-酶联免疫高灵敏检测洛美沙星  

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

作　　者：曹照真[1] 卢圣欣[1] 郗日沫[2] 

机构地区：[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

出　　处：《山东大学学报（理学版）》2011年第11期12-16,共5页Journal of Shandong University(Natural Science)

基　　金：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)

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0.032 ng/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%。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 IC50 value 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%.

关 键 词：抗体 化学发光 酶联免疫 药物残留 洛美沙星 

分 类 号：R917[医药卫生—药物分析学]