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目的:制备抗环丙沙星(CIP)的特异性抗体,以ELISA方法对其进行评价,为进一步研制CIP快速检测试剂盒打基础。方法:以阳离子化牛血清白蛋白(cBSA)为载体合成免疫原CIP-cBSA,免疫新西兰长耳白兔和BALB/c小鼠,同时以天然、阳离子化鸡卵清白蛋白(nOVA、cOVA)为载体合成包被原CIP-nOVA、CIP-cOVA进行抗血清的筛选,以间接(竞争)ELISA法测定抗血清的效价及特异性。结果:本试验获得7种抗血清,其中2种高特异性CIP抗血清,IC50达1μg/L,效价分别为4000和2000(以A450值为1.0时的抗血清稀释倍数表示),两种抗血清与恩诺沙星(ENR)、诺氟沙星(NOR)、氧氟沙星(OFL)存在不同程度的交叉反应,与青霉素(PEN)、卡那霉素(KAN)和庆大霉素(QEN)无交叉。结论:可满足目前中国动物性食品安全检测的需要,为研究组织中CIP残留及开发CIP快速检测试剂盒奠定了基础。
Objective: To prepare anti-ciprofloxacin (CIP) specific antibodies and evaluate them by ELISA, which will lay a foundation for further development of CIP rapid detection kit. Methods: The immunogen CIP-cBSA was synthesized with cationized bovine serum albumin (cBSA) and immunized New Zealand white rabbits and BALB / c mice. The natural and cationized chicken ovary albumin (nOVA, cOVA) The vectors were coated with the original CIP-nOVA and CIP-cOVA for antiserum selection. The antiserum titer and specificity were determined by indirect (competitive) ELISA. Results: Seven kinds of antisera were obtained in this study. Among them, two kinds of high specificity CIP antiserum with IC50 of 1μg / L and titer of 4000 and 2000 respectively (diluted with antiserum at A450 value of 1.0) The antiserum cross-reacted with enrofloxacin (ENR), norfloxacin (NOR) and ofloxacin (OFL) to a certain extent. The antisera reacted with penicillin (PEN), kanamycin No cross (QEN). Conclusion: It can meet the current need for animal food safety testing in China and lay the foundation for the study of CIP residues in tissues and the development of CIP rapid test kit.