Cao Xiaoxiao, Li Shaohua, Chen Liucun, Ding Hongmei, Xu Hua, Huang Yanping, Li Jie, Liu Nongle, Cao Weihong, Zhu Yanjun, Shen Beifen, Shao Ningsheng
Beijing Institute of Basic Medical Sciences, Beijing, China.
Nucleic Acids Res. 2009 Aug;37(14):4621-8. doi: 10.1093/nar/gkp489. Epub 2009 Jun 4.
In this article, a panel of ssDNA aptamers specific to Staphylococcus aureus was obtained by a whole bacterium-based SELEX procedure and applied to probing S. aureus. After several rounds of selection with S. aureus as the target and Streptococcus and S. epidermidis as counter targets, the highly enriched oligonucleic acid pool was sequenced and then grouped under different families on the basis of the homology of the primary sequence and the similarity of the secondary structure. Eleven sequences from different families were selected for further characterization by confocal imaging and flow cytometry analysis. Results showed that five aptamers demonstrated high specificity and affinity to S. aureus individually. The five aptamers recognize different molecular targets by competitive experiment. Combining these five aptamers had a much better effect than the individual aptamer in the recognition of different S. aureus strains. In addition, the combined aptamers can probe single S. aureus in pyogenic fluids. Our work demonstrates that a set of aptamers specific to one bacterium can be used in combination for the identification of the bacterium instead of a single aptamer.
在本文中,通过基于全菌的SELEX程序获得了一组针对金黄色葡萄球菌的单链DNA适配体,并将其应用于探测金黄色葡萄球菌。以金黄色葡萄球菌为靶标,以链球菌和表皮葡萄球菌为反靶标进行了几轮筛选后,对高度富集的寡核苷酸库进行测序,然后根据一级序列的同源性和二级结构的相似性在不同家族下进行分组。从不同家族中选择了11个序列,通过共聚焦成像和流式细胞术分析进行进一步表征。结果表明,5种适配体各自对金黄色葡萄球菌表现出高特异性和亲和力。通过竞争实验,这5种适配体识别不同的分子靶标。在识别不同的金黄色葡萄球菌菌株方面,将这5种适配体组合使用比单独使用一种适配体效果要好得多。此外,组合后的适配体可以探测化脓性液体中的单个金黄色葡萄球菌。我们的工作表明,一组针对一种细菌的适配体可组合用于该细菌的鉴定,而不是使用单个适配体。