可视化蛋白芯片法同时检测牛奶中诺氟沙星和恶喹酸  

Simultaneous detection of norfloxacin and oxolinic acid in milk by visualized microarray

作　　者：李周敏[1] 李心爱[1] 姚开安[1] 许丹科[2] 姚颖 LI Zhou-min;LI Xin-ai;YAO Kai-an;XU Dan-ke;YAO Ying(Jinling College,Nanjing University,Nanjing 210089;State Key Laboratory of Analytical Chemistry for Life Science,School of Chemistry and Chemical Engineering,Nanjing University,Nanjing 210093)

机构地区：[1]南京大学金陵学院,南京210089 [2]南京大学化学化工学院,南京210023

出　　处：《分析试验室》2019年第2期182-186,共5页Chinese Journal of Analysis Laboratory

基　　金：公益性行业(农业)科研专项(201403071);江苏省高等学校自然科学研究项目(18KJD150003)资助

摘　　要：研究了可视化微阵列芯片法同时快速定量检测牛奶中的喹诺酮类抗生素诺氟沙星和恶喹酸的残留量。该法采用了间接竞争免疫法,先在芯片的微孔内以阵列形式固定不同喹诺酮类抗生素的抗原,然后在微孔中加入需要测定的喹诺酮类抗生素的样品与对应抗体以及纳米银标记的二抗,最后加入纳米银催化显色剂,在微孔的芯片上呈现人眼可见的黑点。通过芯片分析仪可获得各黑点的灰度值。通过自动化软件与标准曲线比较后,即可得出喹诺酮类抗生素的残留量,最终达到多种喹诺酮类抗生素的同时定量测定。根据实验结果,标准曲线的相关系数R^2>0. 99,加标回收率在98%～110%之间。检测限(LOD)(3SD)诺氟沙星为0. 1 ng/mL;恶喹酸为0. 2 ng/mL。A method for simultaneous detection of noffloxacin and oxolinic acid in milk by immunoassay without the need for complex or time-consuming pre-treatment procedure was reported.The microarray was prepared by spotting norfloxacin and oxolinie acid antigen onto the microplate.Then the milk samples were diluted with the running buffer and then added into the microplate, followed by the addition of the relevant antibodies as well as silver nanoparticles (AgNPs)labeled secondary antibodies.The measurement principle was based on the competitive immunological reaction and silver enhancement technique.In this case,a visualized array dots as the final signals were further amplified and developed by the developing reagents.The resulting mieroarray could be detected by the microarray scanner.According to the experimental results,the correlation coefficient of the standard curve was measured to R^2>0.99,and the recoveies were 98%-110%.The limits of detection (LOD)(3SD )were estimated to be 0.1 ng/mL (norfloxacin)and 0.2 ng/mL (oxolinic acid), respectively.

关 键 词：可视化 微阵列芯片 诺氟沙星 恶喹酸 银增强 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]