A ratiometric electrochemical DNA-biosensor for detection of miR-141

A sensitive biosensor for the detection of miR-141 has been constructed. The DNA-biosensor is prepared by first immobilizing the thiolated methylene blue-labeled hairpin capture probe (MB-HCP) on two-layer nanocomposite film graphene oxide-chitosan@ polyvinylpyrrolidone-gold nanourchin modified glassy carbon electrode. We used the hematoxylin as an electrochemical auxiliary indicator in the second stage to recognize DNA hybridization via the square wave voltammetry (SWV) responses that record the accumulated hematoxylin on electrode surfaces. The morphology and chemical composition of nanocomposite was characterized using TEM, FE-SEM, and FT-IR techniques. The preparation stages of the DNA-biosensor were screened by electrochemical impedance spectroscopy and cyclic voltammetry. The proposed DNA-biosensor can distinguish miR-141 from a non-complementary and mismatch sequence. A detection limit of 0.94 fM and a linear range of 2.0 -5.0 × 105 fM were obtained using SWV for miR-141 detection. The working potential for methylene blue and hematoxylin was -0.28 and + 0.15 V vs. Ag/AgCl, respectively. The developed biosensor can be successfully used in the early detection of non-small cell lung cancer (NSCLC) by directly measuring miR-141 in human plasma samples. This novel DNA-biosensor is of promise in early sensitive clinical diagnosis of cancers with miR-141 as its biomarker.