【摘要】 基于纳米磁珠标记的免疫层析法,建立快速定量灵敏检测玉米内州萎蔫病菌(Clavibacter michiganensis subsp.nebraskensis,Cmn)的层析试纸条检测方法。用碳二亚胺(EDC)/N-羧基琥珀酰亚胺(NHS)交联法,使玉米内州萎蔫病菌单抗4G12偶联在纳米磁珠表面上,使用喷膜仪将玉米内州萎蔫病菌单抗4H4和羊抗小鼠IgG喷涂于硝酸纤维素膜上,分别作为检测线和质控线,基于双抗夹心法原理建立免疫层析试纸条。通过对检测线上超顺纳米磁珠的磁信号进行检测和结果分析,Cmn抗原最低检测限度为1.0×105 CFU/mL,特异性良好且检测时间缩短为5 min。该方法同时可根据检测线的光学反应进行定性判断,具有操作简便、灵敏度高等优点,并可根据磁信号强度定量检测实际玉米种子样品中Cmn的浓度,为植物病原细菌的快速高效检测提供了一种新型的检测方法。更多还原

【Abstract】 Magnetic nanobead was labeled to establish a rapid and sensitive immunochromatographic strip for the quantitative detection of Clavibacter michiganensis subsp. nebraskensis(Cmn). Magnetic nanobeads were catalyzed and coupled with McAbs 4G12 against for Cmn by ethyl(dimethylaminopropyl) carbodiimide(EDC)/N-Hydroxy succinimide(NHS). The McAbs 4H4 against for Cmn and goat anti-mouse IgG were immobilized on the nitrocellulose membrane as the test line and control line, respectively. The nanobead immunochromatographic strip for detection of Cmn based on double antibody sandwich assay, which arrested the magnic signals. The minimum detection limit of the bacterium was 1.0×105 CFU/mL, and the results came out within 5 min. The results indicate that this method can be used as a rapid,specifi c and sensitive test to detect Cmn in maize seed suspension.更多还原