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| 金黄色葡萄球菌外毒素B特异性适体的筛选及其应用 | |
| 引用本文： | 甘龙杰,兰小鹏,江丽,马立强.金黄色葡萄球菌外毒素B特异性适体的筛选及其应用[J].生物技术通讯,2010,21(2):232-235. |
| 作者姓名： | 甘龙杰 兰小鹏 江丽 马立强 |
| 作者单位： | 1. 福建医科大学,福州总医院临床医学院,福建,福州,350025 2. 福建医科大学,福州总医院临床医学院,福建,福州,350025;南京军区福州总医院,全军医学检验研究所,福建,福州,350025 3. 南京军区福州总医院,全军医学检验研究所,福建,福州,350025 |
| 基金项目： | 全军医学科学技术研究"十一五"计划 |
| 摘 要： | 目的:利用指数富集配基的系统进化(SELEX)技术,筛选能与金黄色葡萄球菌外毒素B(SEB)特异、高亲和力结合的单链DNA(ssDNA)适体,并将该适体应用于患者血清标本的检测。方法:从体外合成的96核苷酸随机ss-DNA文库中,以羧基磁珠作为筛选介质,经逐步PCR扩增、筛选,获得针对SEB的高亲和力、高特异性适体;利用荧光素标记适体测定筛选过程中各轮结合力;利用酶连接适体方法检测适体特异性和结合力。结果:经过13轮筛选,ssDNA文库与SEB的结合百分率从1.1%提高到39.8%,增加了36倍;获得的ssDNA适体(A11)针对SEB的特异性强,与金黄色葡萄球菌表面蛋白A(SPA)结合低,并能初步识别患者血清。结论:利用SELEX技术筛选获得了特异结合SEB的高亲和力的ssDNA适体,为金黄色葡萄球菌的临床诊断与治疗奠定了基础。 |
| 关 键 词： | 指数富集的配体系统进化技术 金黄色葡萄球菌外毒素B 适体 |
Selection and Application of DNA Aptamers Specifically to Staphylococcal aureus Enterotoxin B |
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| GAN Long-Jie,LAN Xiao-Peng,JIANG li,MA Li-Qiang.Selection and Application of DNA Aptamers Specifically to Staphylococcal aureus Enterotoxin B[J].Letters in Biotechnology,2010,21(2):232-235. | |
| Authors: | GAN Long-Jie LAN Xiao-Peng JIANG li MA Li-Qiang |
| Institution: | GAN Long-Jie1,LAN Xiao-Peng1,2,JIANG li1,MA Li-Qiang2 1.Clinical Medicine School,Fuzhou General Hospital,Fujian Medical University,Fuzhou 350025,2.Institue of Laboratory Medicine,Fuzhou General Hospital of PLA,China |
| Abstract: | Objective:The systematic evolution of ligands by exponential enrichment (SELEX) was applied for screening single strand DNA(ssDNA) aptamers which specifically bind to Staphylococcal aureus enterotoxin B(SEB),and was used in clinical detection.Methods:A 96 nt random ssDNA library was designed and synthesized in vitro,and the dynabeads carboxylic acid was as medium in the SELEX,the ssDNA aptamers specifically binding to SEB were obtained through PCR amplification and selection.The aptamer exhibited binding ab... |
| Keywords: | systematic evolution of ligands by exponential enrichment Staphylococcal aureus enterotoxin B aptamer |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |