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当前位置: 首页 > 文章 > 诺氟沙星免疫学检测方法的建立及优化 食品科学 2011,32 (21) 148-151
Position: Home > Articles > Development and Optimization of Immunoassay Method for Detection of Norfloxacin Residues FOOD SCIENCE 2011,32 (21) 148-151

诺氟沙星免疫学检测方法的建立及优化

作  者：
姜金庆;李广领;王自良;刘兴友;杨雪峰
单  位：
河南科技学院动物科学学院;动物疫病和残留物防控河南省高校工程技术研究中心
关键词：
诺氟沙星;氟喹诺酮;间接竞争ELISA;多残留检测
摘  要：
采用优化的碳二亚胺法制备诺氟沙星(NOR)人工抗原,免疫新西兰大白兔制备多克隆抗体,建立间接竞争ELISA标准曲线。对NOR的线性检测范围为0.07～31.8ng/mL,半数抑制质量浓度(IC50)和最低检测限(LOD)分别为1.5ng/mL和0.04ng/mL。检测缓冲液中pH值和磷酸根离子浓度的最佳参数分别为7.4和10mmol/L;对甲醇和NaOH溶液(30mmol/L)的最佳调整体积分数分别为30%和10%。抗体具有广谱特异性,与环丙沙星(83.3%)、培氟沙星(78.9%)、依诺沙星(68.2%)、恩诺沙星(51.7%)和洛美沙星(41.7%)有较高的交叉反应率,可用于氟喹诺酮类药物(FQs)多残留检测方法研究。
译  名：
Development and Optimization of Immunoassay Method for Detection of Norfloxacin Residues
作  者：
JIANG Jin-qing1,2,LI Guang-ling2,WANG Zi-liang2,,LIU Xing-you2,YANG Xue-feng1,HUANG Hua-guo1(1.College of Animal Science,Henan Institute of Science and Technology,Xinxiang 453003,China;2.Henan Higher Education Engineering Technology Research Center for Animal Diseases Control and Residues Supervison,Xinxiang 453003,China)
关键词：
norfloxacin(NOR);fluoroquinolone(FQs);indirect competitive ELISA;multi-residue detection
摘  要：
Modified EDC method was employed to synthesize norfloxacin(NOR) artificial antigen and New Zealand rabbits were immunized for anti-NOR polyclonal antibody(pAb) production.An indirect competitive ELISA standard curve was established.The developed assay was sensitive and had a linear range from 0.07 to 31.8 ng/mL(R2 = 0.9627),with IC50 and LOD of respectively 1.5 ng/mL and 0.04 ng/mL.The optimal pH and phosphate ion concentration in assay buffer were determined to be 7.4 mmol/L and 10 mmol/L,respectively,and the concentration adjustments for methanol and NaOH(30 mmol/L) in dilute solutions did not exceed 30% and 10%,respectively.The produced pAb had broad-spectrum specificity and exhibited a high cross-reactivity with ciprofloxacin(83.3%),pefloxacin(78.9%),enoxacin(68.2%),enrofloxacin(51.7%) and lomefloxacin(41.7%).The results obtained suggest that the established icELISA method be used for the multi-determination of fluoroquinolone residues.

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