直接竞争ELISA法快速测定水产品中6种氟喹诺酮类药物

Rapid determination of six fluoroquinolones in aquatic product by direct competitive enzyme-linked immunosorbent assay

采用过碘酸钠法将抗环丙沙星单克隆抗体（MAb-CIP）与辣根过氧化物酶（HRP）偶联制备酶标抗体MAbCIP-HRP,建立了检测水产品中6种氟喹诺酮类药物残留的直接竞争酶联免疫吸附分析方法（dcELISA）,考察了包被原浓度、竞争反应时间和有机溶剂等因素对方法灵敏度的影响。结果表明：在优化的反应条件下,所建立的dcELISA针对环丙沙星、恩诺沙星、诺氟沙星、培氟沙星、沙拉沙星和双氟沙星6种氟喹诺酮类药物的检测限（LOD）均不超过0.5ng/mL,线性范围（IC20-IC80）在1.0-12.1ng/mL之间;对虾、鳗鲡和鲫鱼三种水产样品中添加5.0、10.0和20μg/kg时,加标回收率为70.4%-104.1%,相对标准偏差为5.0%~14.7%;本方法可用于水产品中氟喹诺酮类（FQs）药物多残留的快速测定。

In this study,a direct competitive enzyme-linked immunosorbent assay（dcELISA） was developed for rapid determination six fluoroquinolones in aquatic product. Horseradish peroxidase-labelled ciprofloxacin monoclonal antibody（ MAb-CIP-HRP） was synthesized by NaIO4method. The effect of coating antigen concentration,competitive reaction time and organic solvents on the sensitivity of dcELISA were investigated.Under the optimized assay conditions,the dcELISA showed the limit of detection below 0.5ng/mL for ciprofloxacin,enrofloxacin,norfloxacin,pefloxacin,sarafloxacin and difloxacin,and the linear range（IC20~IC80） of 1.0 to 12.1ng/mL.Recoveries from spiked shrimp,eel and crucian at levels of 5.0,10 and 20μg/kg were in the range of 70.4%~104.1%,with relative standard deviation ranging from 5.0% to 14.7%. Results indicated that the established dcELISA was suitable for multi-analytes rapid determination of fluoroquinolones residues in aquatic product.