Journal
ACS COMBINATORIAL SCIENCE
Volume 18, Issue 6, Pages 292-301Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acscombsci.5b00123
Keywords
whole cell SELEX; DNA aptamer; Salmonella Typhimurium; fluorescence; gold nanoparticle
Funding
- Department of Science and Technology (DST
- New Delhi, India)
- INSPIRE program
Ask authors/readers for more resources
This study describes the selection of single stranded DNA (ssDNA) aptamers against Salmonella enterica serovar Typhimurium using a modified whole cell systematic evolution of ligands by exponential enrichment (whole cell SELEX). For evolving specific aptamers, ten rounds of selection to live Salmonella cells, alternating with negative selection against a cocktail of related pathogens, were performed. The resulting highly enriched oligonucleotide pools were sequenced and clustered into eight groups based on primary sequence homology and predicted secondary structure similarity. Fifteen sequences from different groups were selected for further characterization. The binding affinity and specificity of aptamers were determined by fluorescence binding assays. Aptamers (SAL 28, SAL 11, and SAL 26) with dissociation constants of 195 46, 184 43, and 123 23 nM were used to develop a nanogold-based colorimetric detection method and a sedimentation assay. The former showed a better sensitivity limit of 102 CFU/mL using aptamer SAL 26. This approach should enable further refinement of diagnostic methods for the detection of Salmonella enterica serovar Typhimurium and of other microbial pathogens.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available