Design of Electrochemical Nanobiosensors in the Application Kit Type for the Determination of DNA Sequences
Project no. 294 · Status: Kapanmış · 2018
Summary
According to World Health Organization data, antibiotic resistance causes the death of 700.000 people every year. Unless a solution is provided; it is estimated that the number of people who died due to antibiotic resistance by 2050 will increase up to 10 million. In this study, gene regions that transmit the resistance mechanism of carbapenemase enzyme which responsible for more than 50% of antibiotic resistance deaths were investigated. In this study; for the determination of OXA-48 and VIM gene regions which are the carbapenemase resistance genes, a ready-to-use kit type electrochemical biosensor design have been made with pencil graphite electrode. In order to convert the nanobiosensor to the kit system developed for the antibiotic resistance analysis, the synthetic probe sequences immobilize on to electrod surfaces which modified with nanomaterials, and then this electrodes kept under appropriate storage conditions. In studies involving with synthetic DNA sequences, determination of target sequences was carried out for 30 minutes by enzyme mediated hybridization with a kit-type biosensor. In addition, symmetric and asymmetric polymerase chain reaction (PCR) products of OXA-48 and VIM were determined by using ready-to-use kit type biosensor. It was found that the developed kit-type biosensor can perform gene analysis even after 150 days of kit preparation. Due to the effect of nanomaterial modification, sensitive determination was achieved and the limit of detection was calculated as 2.50 picomole/50µL. As a result, DNA-based antibiotic resistance gene analysis was performed with kit type nanobiosensor and a new alternative method was introduced to the classical methods. Keywords: Antibiotic resistance, kit, electrochemical DNA nanobiosensor, carbon nanotube, polymerase chain reaction (PCR), scanning electron microscopy (SEM).