کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2090030 1081467 2013 5 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Identification of Salmonella Typhimurium-specific DNA aptamers developed using whole-cell SELEX and FACS analysis
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی بیوتکنولوژی یا زیست‌فناوری
پیش نمایش صفحه اول مقاله
Identification of Salmonella Typhimurium-specific DNA aptamers developed using whole-cell SELEX and FACS analysis
چکیده انگلیسی


• We developed aptamers which has specific binding capabilities to S. Typhimurium.
• We modified and adopted the whole-cell SELEX for the development of ssDNA aptamers.
• We examined those binding specificities by FACS analysis.
• Total 12 aptamer candidates were selected and sequenced after SELEX and FACS.
• Aptamer C4 was selected, as to be had the highest binding affinity and selectivity.

Conventional methods for detection of infective organisms, such as Salmonella, are complicated and require multiple steps, and the need for rapid detection has increased. Biosensors show great potential for rapid detection of pathogens. In turn, aptamers have great potential for biosensor assay development, given their small size, ease of synthesis and labeling, lack of immunogenicity, a lower cost of production than antibodies, and high target specificity. In this study, ssDNA aptamers specific to Salmonella Typhimurium were obtained by a whole bacterium-based systematic evolution of ligands by exponential enrichment (SELEX) procedure and applied to probing S. Typhimurium. After 10 rounds of selection with S. Typhimurium as the target and Salmonella Enteritidis, Escherichia coli and Staphylococcus aureus as counter targets, the highly enriched oligonucleic acid pool was sorted using flow cytometry. In total, 12 aptamer candidates from different families were sequenced and grouped. Fluorescent analysis demonstrated that aptamer C4 had particularly high binding affinity and selectivity; this aptamer was then further characterized.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Microbiological Methods - Volume 95, Issue 2, November 2013, Pages 162–166
نویسندگان
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