玉米细菌性枯萎病菌纳米磁标记免疫层析检测试剂与试纸条  

Development of Magnetic Nanobead-based Immunochromatographic Detection Reagents and Strips for Pantoea Stewartii Subsp. Stewartin

作　　者：杨金玲[1] 黄大亮 李莉[1] 刘彩虹[1] 陈勇 邹明强[1,4] 陈艳[1,4] 马吉湘[1] 

机构地区：[1]中国检验检疫科学研究院,北京100123 [2]大连出入境检验检疫局,辽宁大连116001 [3]山西出入境检验检疫局,山西太原030024 [4]中检国研(北京)科技有限公司,北京100123

出　　处：《化学试剂》2015年第6期519-523,共5页Chemical Reagents

基　　金：国家重大科学仪器设备开发专项资助项目(2012YQ090197);"十二五"国家科技支撑计划课题(2012BAK-26B04);质检公益性行业科研专项资助项目(201010022)

摘　　要：分别以粒径为50和100 nm的羧基化纳米磁珠,通过碳二亚胺(EDC)/N-羟基琥珀酰亚胺(NHS)交联法使磁珠与抗体偶联,制备出可特异性识别玉米细菌性枯萎病菌的两种纳米免疫磁珠试剂。基于双抗体夹心法,经实验条件优化,选用100 nm的羧基化纳米磁珠为磁标记探针,组装了玉米细菌性枯萎病菌磁标记免疫层析检测试纸条。该方法用于玉米种子中玉米细菌性枯萎病菌的检测,检出限为1.0×106CFU/m L,15 min内可通过肉眼判读结果,也可用超顺磁共振检测仪实现量化检测,具有简便、快速、现场化检测的优势。Magnetic nanobead-based immunochromatographic detection reagents for Pantoea stewartii subsp, stewartii were pre- pared,using respectively two kinds of sizes （50 nm and 100 nm） carboxylated magnetic nano-beads by activating reaction with ethyl （dimethylaminopropyl） carbodiimide （EDC）/N-hydroxy succinimide （NHS） as activators. Based on double antibody sand- wich assay,the magnetic immunochromatography strip for detection of Pantoea stewartii subsp, stewartii was optimized and finally assembled with the 100 nm magnetic nano-beads as magnetically labeled probes. The bacteria was detected Within 15 min by the naked eyes or using magnetic assay reader to achieve quantitative detection,with a detection limit of 1.0 × 10^6 CFU/mL. It is sim- ple,rapid and easily available for on-site screening for Pantoea stewartii subsp, stewartii in corn seeds.

关 键 词：玉米细菌性枯萎病菌 纳米磁珠 免疫层析检测 试剂 试纸条 

分 类 号：Q7[生物学—分子生物学] S41[农业科学—植物保护]