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Selection of DNA aptamers to Streptococcus pneumonia and fabrication of graphene oxide based fluorescent assay

Bayraç, Abdullah Tahir, Donmez, Sultan Ilayda
Analytical biochemistry 2018 v.556 pp. 91-98
DNA, Streptococcus pneumoniae, antibiotic resistance, bacteria, binding capacity, children, elderly, flow cytometry, fluorescence, fluorescence emission spectroscopy, graphene oxide, nucleotide aptamers, patients, pneumonia, public health, systematic evolution of ligands by exponential enrichment
Pneumococci are one of the leading causes of infections throughout the world causing problems mainly in children, elderly, and immune-deficient patients. In recent years antibiotic resistant Streptococcus pneumoniae strains become widespread. Therefore simple, rapid, and specific detection methods are needed for public health. In this study, DNA aptamer probes against S. pneumoniae were selected using bacterial Systematic Evolution of Ligands by Exponential Enrichment (SELEX) and these probes were integrated in to a graphene oxide (GO) based fluorescent assay. Among the tested aptamers three candidates Lyd-1, Lyd-2 and Lyd-3 showed Kd values of 844.7 ± 123.6, 1984.8 ± 347.5, and 661.8 ± 111.3 nM, respectively. These candidates showed binding affinity to S. pneumoniae and no specific binding to the bacteria used in negative selection. The binding of aptamers were showed by fluorescence spectroscopy and flow cytometry. GO based label-free fluorescent assay developed using Lyd-3 aptamer had a unique detection limit of 15 cfu mL⁻¹. Thus we believe that the selected aptamers and fabricated GO based assay has potential to be used in the detection of S. pneumoniae. Selected aptamers selectively bind to S. pneumonia with anti-pneumococcal potential and holds great potential to be used as molecular probes for identifying and targeting.