Anaplasma Phagocytophilum’un Tayinine Yönelik Elektrokimyasal DNA Sensörü Geliştirilmesi

Bu çalışma, oldukça geniş bir konakçı gurubunda enfeksiyona yol açan bir patojen olan Anaplasma Phagocytophilum dizi seçimli DNA hibridizasyonunun voltametrik yöntemle tayin edildiği tek kullanımlık elektrokimyasal DNA biyosensörü geliştirilmesine yöneliktir. Herhangi bir etiketlemenin yapılmadığı bu elektrokimyasal çalışmanın esasını DNA hibridizasyonu sonrasında Guanin bazının yükseltgenme sinyalinin ölçülmesi ile bir dubleks oluşumunun izlenmesi oluşturur. Söz konusu biyosensör tasarımı inozinmodifiye (guanin içermeyen) probun, kalem grafit elektrota (PGE) immobilize edilmesi ve diferansiyel puls voltametrisi (DPV) ile guanin yükseltgenme sinyalinin ölçülerek, dubleks oluşumu tayinini içerir. Bu çalışmanın ilk aşamasında, Anaplasma phagocytophilum temsil eden prob dizisi, aktive edilmiş kalem grafit elektrot (PGE) yüzeyine yaş adsorbsiyon yöntemi ile immobilize edilmiş ve daha sonra prob ve hedefi arasındaki hibridizasyonun varlığı 1000 mV'da gözlenen guanin yükseltgenme sinyali ile tespit edilmiştir

Development of Electrochemical DNA Sensor for The Determination of Anaplasma Phagocytophilum

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 voltammetry 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). 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

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