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

Cao X, Li S, Chen L, Ding H, Xu H, Huang Y, Li J, Liu N, Cao W, Zhu Y, Shen B, Shao N - Nucleic Acids Res. (2009)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Beijing Institute of Basic Medical Sciences, Beijing, China.

ABSTRACT
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.

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Related in: MedlinePlus

The monitoring of enrichment of selection process by binding assay of radiolabeld pools with S. aureus. The number 0 represents the naive library and the numbers 1–7 represents the first to seventh round selected pool, respectively. Twenty-five picomoles of each radiolabeled pool were incubated with 107S. aureus 8325-4 for 45 min. The binding capacity was detected by scintillation counting.
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Figure 2: The monitoring of enrichment of selection process by binding assay of radiolabeld pools with S. aureus. The number 0 represents the naive library and the numbers 1–7 represents the first to seventh round selected pool, respectively. Twenty-five picomoles of each radiolabeled pool were incubated with 107S. aureus 8325-4 for 45 min. The binding capacity was detected by scintillation counting.

Mentions: The enrichment of selection pools were monitored by the binding assays of enriched pools. The results showed directly that fifth round pool was enriched best with radioactivity of about 100-fold excess of the naive ssDNA library (Figure 2). The sixth and seventh round showed a gradually decreased enrichment, which may be a result of loss of structured specific aptamers through PCR amplification.Figure 2.


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

Cao X, Li S, Chen L, Ding H, Xu H, Huang Y, Li J, Liu N, Cao W, Zhu Y, Shen B, Shao N - Nucleic Acids Res. (2009)

The monitoring of enrichment of selection process by binding assay of radiolabeld pools with S. aureus. The number 0 represents the naive library and the numbers 1–7 represents the first to seventh round selected pool, respectively. Twenty-five picomoles of each radiolabeled pool were incubated with 107S. aureus 8325-4 for 45 min. The binding capacity was detected by scintillation counting.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2724295&req=5

Figure 2: The monitoring of enrichment of selection process by binding assay of radiolabeld pools with S. aureus. The number 0 represents the naive library and the numbers 1–7 represents the first to seventh round selected pool, respectively. Twenty-five picomoles of each radiolabeled pool were incubated with 107S. aureus 8325-4 for 45 min. The binding capacity was detected by scintillation counting.
Mentions: The enrichment of selection pools were monitored by the binding assays of enriched pools. The results showed directly that fifth round pool was enriched best with radioactivity of about 100-fold excess of the naive ssDNA library (Figure 2). The sixth and seventh round showed a gradually decreased enrichment, which may be a result of loss of structured specific aptamers through PCR amplification.Figure 2.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Beijing Institute of Basic Medical Sciences, Beijing, China.

ABSTRACT
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.

Show MeSH
Related in: MedlinePlus