Combining use of a panel of ssDNA aptamers in the detection of Staphylococcus aureus

被引:283
作者
Cao, Xiaoxiao [1 ]
Li, Shaohua [1 ]
Chen, Liucun [1 ]
Ding, Hongmei [1 ]
Xu, Hua [1 ]
Huang, Yanping [2 ]
Li, Jie [1 ]
Liu, Nongle [1 ]
Cao, Weihong [3 ]
Zhu, Yanjun [4 ]
Shen, Beifen [1 ]
Shao, Ningsheng [1 ]
机构
[1] Beijing Inst Basic Med Sci, Beijing, Peoples R China
[2] Gen Hosp Chinese Peoples Armed Police Forces, Beijing, Peoples R China
[3] Peoples Liberat Army Gen Hosp, Clin Div 304, Beijing, Peoples R China
[4] Gen Hosp PLA Air Force, Beijing, Peoples R China
关键词
IN-VITRO SELECTION; EXPONENTIAL ENRICHMENT; SYSTEMATIC EVOLUTION; DNA APTAMERS; LIGANDS; CELLS; BIND; INFECTIONS; VIRULENCE; CANCER;
D O I
10.1093/nar/gkp489
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:4621 / 4628
页数:8
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