Impedimetric genosensor for miRNA-34a detection in cell lysates using polypyrrole
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This work reports the development of simple, practical, cost effective and label free genosensor prepared by electropolymerization of polypyrrole on pencil graphite electrode (PGE) for the determination of miRNA-34a from total RNA extracted from breast cancer cell lysate. The electrochemical entrapment of the probe (antimiRNA-34a) into polypyrrole (PPy) was carried out by electropolymerization using cyclic voltammetry method with a scan rate of 25 mV.s−1 versus Ag/AgCl. The electrochemical detection of the hybridization between the doped probe antimiRNA-34a with its complementary target, miRNA-34a was monitored by electrochemical impedance spectroscopy (EIS) by comparison of charge transfer resistance (Rct) values before and after hybridization. The established biosensor can detect miRNA-34a down to 0.2 μg.mL−1 (which correspond to 2 pmol/100 μL) with a linear range of 5–80 μg.mL−1 and discriminate target miRNA from other non-complementary sequences (miRNA-21, miRNA-122, and miRNA-192) with a high selectivity. The genosensor shows a better performance in analysis of human breast cancer cells samples (MCF-7) suggesting the real-time usability of the genosensor.
KeywordsElectrochemical impedance spectroscopy miRNA-34a Human breast cancer cells Electropolymerization Polypyrrole Pencil graphite electrode
The authors would like to acknowledge Professor Mehmet Özsöz and his team of researchers at Gediz University, Turkey, for the fruitful discussion and partial support of this work.
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