Development of Electrochemical DNA Sensor for The Determination of Anaplasma Phagocytophilum
Journal Title: Cumhuriyet Science journal - Year 2017, Vol 38, Issue 4
Abstract
This study was intended to enhance the disposable electrochemical DNA biosensor through which Anaplasma phagocytophilum, a pathogen causing infection in a considerably wide host group, sequence selective DNA hybridization was detected with a voltametric method. Monitoring the formation of a duplex by measuring the oxidation signal of guanine base after DNA hybridization forms the basis of this study in which no labeling was done. The biosensor design contained immobilizing inosine modified (guanine-free) probe to the pencil graphite electrode (PGE) and detecting duplex formation by measuring guanine oxidation signal with differential pulse voltammetry (DPV). In the first stage of this study, probe sequence representing Anaplasma phagocytophilum was immobilized to the activated surface of pencil graphite electrode (PGE) by wet adsorbtion method, and then the hybridization between probe and its target was confirmed with guanine oxidation signal observed at 1000 mV. In the optimization study, it was observed that probe concentration was 25 µg/mL, immobilization time was 6 minutes; for the hybridization, target concentration was 40 µg/mL and hybridization time was 10 minutes. Selectivity of biochemical biosensor developed was tested by using mismatch and non complementary target sequences. It was also confirmed that DNA hybridization was carried out with impedimetric measurements by using electrochemical impedance method under the ferri/ferro cyanide redox system. To estimate the detection limit (DL), target sequences whose concentrations varies between 0, 78 µM and 3,90 µM were used, and it was found that the detection limit was 0,244 µM.
Authors and Affiliations
Adil ELİK, Gültekin GÖKÇE, Çağdaş CEYLAN
Keywords
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- EP ID EP488460
- DOI 10.17776/csj.349331
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