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Nucleic Acids Research Advance Access published online on June 4, 2009
Nucleic Acids Research, doi:10.1093/nar/gkp489
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© 2009 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Molecular Biology

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

Xiaoxiao Cao1, Shaohua Li1, Liucun Chen1, Hongmei Ding1, Hua Xu1, Yanping Huang2, Jie Li1, Nongle Liu1, Weihong Cao3, Yanjun Zhu4, Beifen Shen1 and Ningsheng Shao1,*

1Beijing Institute of Basic Medical Sciences, 2General Hospital of Chinese People's Armed Police Forces, 3304th Clinical Division of PLA General Hospital and 4General Hospital of PLA Air Force, Beijing, China

*To whom correspondence should be addressed. Tel: +86 10 68163140; Fax: +86 10 68163140; Email: shaonsh{at}hotmail.com

Received January 22, 2009. Revised April 27, 2009. Accepted May 19, 2009.

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.

The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.


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