Development of enrofloxacin ELISA using a monoclonal antibody tolerating an organic solvent with broad cross-reactivity to other newquinolones
Development of enrofloxacin ELISA using a monoclonal antibody tolerating an organic solvent with broad cross-reactivity to other newquinolones
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DOI：
10.1080/09540100701763365
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发表时间：
2007-01-01
影响因子：
3
通讯作者：
Nakazawa, Hiroyuki
中科院分区：
文献类型：
--
作者：
Kato, Mihoko;Ihara, Yoshiharu;Nakazawa, Hiroyuki
抗菌试剂，尤其是喹诺酮类，广泛应用于动物和人类，由于其在食物中的残留污染物，已对人类健康造成严重问题。为了同时筛选不同种类的新型喹诺酮类药物，一种灵敏且特异的酶联免疫吸附测定法（ELISA）已被研发出来。选择抗恩诺沙星单克隆抗体是因为它在有机溶剂中能与结构相关的新型喹诺酮类药物发生反应。该抗体在对照值抑制率为50%（IC50）时，与诺氟沙星、环丙沙星及其他新型喹诺酮类药物有100%的交叉反应，但与硝呋拉宗、磺胺地索辛无交叉反应。当以恩诺沙星作为校准物时，这种ELISA的最低检测限为0.7纳克/毫升（十亿分之一）。鳗鱼提取物中添加恩诺沙星后，在10、50、100纳克/毫升浓度下的平均回收率分别为98%、102%和91%。所提出的ELISA是一种对生物和环境样本中各种新型喹诺酮类药物进行实际微量定量的有用方法。
Antibacterial reagents, especially quinolones, are widely used in animals and humans, and have caused serious problems to human health because of their residual contaminants in food. In order to screen for different kinds of newquinolones at the same time, a sensitive and specific enzyme-linked immunoassay (ELISA) has been developed. The anti-enrofloxacin monoclonal antibody was selected because of its ability to react with structurally related newquinolones in organic solvent. The antibody has 100% cross-reactivity with norfloxacin, ciprofloxacin and other newquinolones at 50% inhibition of control values IC50, but not with nitroflazone, sulphadimethoxine. The lowest detection limit of this ELISA was 0.7 ng/ml (ppb) when enrofloxacin was used as the calibrator. Eel extracts were spiked with enrofloxacin and the average recoveries at 10, 50, 100 ng/ml were 98, 102 and 91%, respectively. The proposed ELISA is a useful method for the practical microquantitation of various newquinolones in biological and environmental specimens.