SELEX技术筛选耐甲氧西林金黄色葡萄球菌适配子的研究  

In vitro selection of aptamers against methicillin-resistant Staphylococcus aureus by SELEX

作　　者：汪江波[1] 李赞东[1] 赵延斌[1] 侯德凤[2] 廖玲[3] 李艳红[4] 毛伟丽[5] 

机构地区：[1]广西壮族自治区南溪山医院药剂科,广西桂林541002 [2]广西壮族自治区南溪山医院检验科,广西桂林541002 [3]广西壮族自治区南溪山医院科教科,广西桂林541002 [4]广西壮族自治区南溪山医院医保科,广西桂林541002 [5]广州医学院,广东广州510182

出　　处：《中华医院感染学杂志》2013年第13期3041-3043,共3页Chinese Journal of Nosocomiology

基　　金：广西壮族自治区卫生厅资助项目(Z2010075)

摘　　要：目的利用指数富集的配体进化技术(SELEX)筛选耐甲氧西林金黄色葡萄球菌(MRSA)的适配子,并研究适配子AM6与MRSA结合的特异性,为研发MRSA的特异性诊断试剂和靶向治疗药物打下基础。方法体外构建81个碱基的随机单链DNA(ssDNA)文库;通过反复的SELEX筛选获得MRSA的适配子并进行一级结构分析;利用荧光显微镜和PCR扩增法鉴定AM6与MRSA结合的特异性。结果经过20轮环筛选,获得了25个与预期大小相同的ssDNA序列,它们之间没有共同的保守序列;AM6仅与MRSA结合,而不与甲氧西林敏感葡萄球菌(MSSA)、耐甲氧西林凝固酶阴性葡萄球菌(MRSCN)、甲氧西林敏感凝固酶阴性葡萄球菌(MSSCN)、大肠埃希菌结合。结论利用SELEX技术成功地筛选出MRSA的适配子,其中适配子AM6能特异性结合MRSA。OBJECTIVE To obtain aptamers of methicillin-resistant Staphylococcus aureus（MRSA）by SELEX and observe the binding specificity of aptamer AM6to MRSA so as to lay foundation for developing specific diagnostic reagents and targeted therapies on MRSA.METHODS An 81nt primary random ssDNA library was synthesized in vitro.Aptamers of MRSA were obtained by iterative SELEX procedures,and then their primary structures were analyzed.The specificity of AM6＇s binding to MRSA was observed by fluorescence microscope and PCR.RESULTS After 20rounds of selection,25ssDNA sequences the same as the expected size were obtained,which had no common conserved sequence.AM6did not bind to methicillin-sensitive Staphylococcus aureus（MSSA）,methicillin-resistant coagulase-negative Staphylococcus（MRCNS）,or methicillin-sensitive coagulase-negative Staphylococcus（MSCNS）,and Escherichia coli,but it bound to MRSA.CONCLUSIONThe aptamers of MRSA can be successfully obtained by SELEX,and AM6can bind to MRSA specifically.

关 键 词：指数富集的配体进化技术 耐甲氧西林金黄色葡萄球菌 适配子 

分 类 号：R446[医药卫生—诊断学]