Isolation of an Aptamer that Binds Specifically to E. coli.
Isolation of an Aptamer that Binds Specifically to E. coli.
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DOI：
10.1371/journal.pone.0153637
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Cardoso J
中科院分区：
文献类型：
--
作者：
Marton S;Cleto F;Krieger MA;Cardoso J
大肠杆菌是一种在动物肠道植物群中普遍存在的细菌物种，尽管致病性变体会引起重大的公共卫生问题。适体是短的寡核苷酸，其以高亲和力和特异性结合到靶标，并且在诊断和治疗中具有巨大的应用潜力。我们使用基于细胞的指数富集配体系统进化（cell-SELEX）技术分离了四种与大肠杆菌强结合的单链DNA适体。coli细胞（ATCC通用菌株25922），Kd值在纳摩尔范围内。荧光标记的适体标记E.大肠杆菌细胞，如通过荧光显微镜观察。对12种不同的细菌物种进行的特异性测试表明，其中一种称为P12-31的适体对E.杆菌重要的是，该适体结合脑膜炎/脓毒症相关的E.大肠杆菌（MNEC）临床分离株，是第一个适体描述的潜在用于诊断MNEC传播的病理。
Escherichia coli is a bacterial species found ubiquitously in the intestinal flora of animals, although pathogenic variants cause major public health problems. Aptamers are short oligonucleotides that bind to targets with high affinity and specificity, and have great potential for use in diagnostics and therapy. We used cell-based Systematic Evolution of Ligands by EXponential enrichment (cell-SELEX) to isolate four single stranded DNA (ssDNA) aptamers that bind strongly to E. coli cells (ATCC generic strain 25922), with Kd values in the nanomolar range. Fluorescently labeled aptamers label the surface of E. coli cells, as viewed by fluorescent microscopy. Specificity tests with twelve different bacterial species showed that one of the aptamers–called P12-31—is highly specific for E. coli. Importantly, this aptamer binds to Meningitis/sepsis associated E. coli (MNEC) clinical isolates, and is the first aptamer described with potential for use in the diagnosis of MNEC-borne pathologies.