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

被引:38
作者
Bayrac, Abdullah Tahir [1 ]
Donmez, Sultan Ilayda [1 ]
机构
[1] Karamanoglu Mehmetbey Univ, Dept Bioengn, Yunus Emre Campus, TR-70100 Karaman, Turkey
关键词
Aptamer; pneumoniae; SELEX; Pathogen biosensor; Biofilm; IN-VITRO SELECTION; BIOFILM FORMATION; SSDNA APTAMERS; SELEX TECHNIQUE; CELL-SELEX; IDENTIFICATION; BACTERIAL; MULTIPLE; MICROBIOLOGY; STRAINS;
D O I
10.1016/j.ab.2018.06.024
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 K-d 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.
引用
收藏
页码:91 / 98
页数:8
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