纤维素磁性微球在病原微生物检测中的应用研究  

APPLICATION RESEARCH OF MAGNETIC CELLULOSE MICROSPHERES IN THE DETECTION OF PATHOGENIC MICROORGANISIMS

作　　者：白芳[1] 张奇[1] 王龙[2] 张玺[1] 孙丹[3] 白钢[1,2] 

机构地区：[1]南开大学药学院,天津300071 [2]南开大学生命科学学院,天津300071 [3]南开大学学报编辑部,天津300071

出　　处：《离子交换与吸附》2010年第5期431-438,共8页Ion Exchange and Adsorption

基　　金：天津市应用基础与前沿技术研究计划(08JCYBJC04400)

摘　　要：采用反向悬浮包埋技术和高速匀浆法制备了不同粒径的纤维素磁性微球,扫描电镜和铁离子分析结果表明,最佳粒径为5.82μm。后采用自行构建并表达的融合蛋白CBD-ProA和沙门氏菌多价抗体对最佳粒径的纤维素磁性微球进行活化,获得免疫磁性微球,其抗体偶联量为186.8mg/mL。最后采用致敏的免疫磁性微球分别对污染沙门氏菌的饮用水、牛奶和花生酱等样品进行免疫磁性捕获及分子鉴定,其检测限均可达10cfu/100mL。本研究开发的免疫磁性捕获技术,具有微球活化速度快、工艺简单、使用便捷等优势,同时,建立的免疫磁性捕获-PCR的方法与国标法相比,检测时间缩短48h,在食品安全、环境检测等方面具有实用价值。Six different particle sizes of the magnetic cellulose microspheres（MCMS）were prepared by reverse embedding technology and high-speed suspension homogenates,and the particle size of 5.82μm was best by scanning electron microscopy and the iron ion analysis.Then anti-Salmonella multi-serum was fixed on surface of the best size MCMS with the expressed the fusion protein CBD-ProA,the amount of antibody-coupled was 186.8mg/mL.The sensitized MCMS was used to capture Salmonella of drinking water,milk and peanut butter,respectively.After culturing on LB,the detection limit could reach 10cfu/100mL.Then molecular identification was processed to verify the results of the previous.The method of immunomagnetic capture-PCR was faster than national standard method,and 2 days could be saved.The results showed that the method could be used in several fields including food safety testing and environmental detection.

关 键 词：沙门氏菌 免疫磁性捕获 CBD-ProA 

分 类 号：TS207.4[轻工技术与工程—食品科学]