RESUMO
Due to the late detection of stomach cancer, this cancer usually causes high mortality. The development of an electrochemical genosensor to measure microRNA 106b (miR-106b), as a gastric cancer biomarker, is the aim of this effort. In this regard, first, 1,3,5-benzenetricarboxylate (BTC) metal-organic frameworks (Zn-BTC MOF) were self-assembled on the glassy carbon electrode and then the probe (ssDNA) was immobilized on it. The morphology Zn-BTC MOF was characterized by SEM, FT-IR, Raman and X-Ray techniques. Zn-BTC MOF as a biosensor substrate has strong interaction with ssDNA. Quantitative measurement of miR-106b was performed by electrochemical impedance spectroscopy (EIS). To perform this measurement, the difference of the charge transfer resistances (ΔRct) of Nyquist plots of the ssDNA probe modified electrode before and after hybridization with miR-106b was obtained and used as an analytical signal. Using the suggested genosensor, it is possible to measure miR-106b in the concentration range of 1.0 fM to 1.0 µM with a detection limit of 0.65 fM under optimal conditions. Moreover, at the genosensor surface, miR-106b can be detected from a non-complementary and a single base mismatch sequence. Also, the genosensor was used to assess miR-106b in a human serum sample and obtained satisfactory results.
Assuntos
Técnicas Biossensoriais , MicroRNAs , Neoplasias Gástricas , Humanos , Biomarcadores Tumorais/genética , Neoplasias Gástricas/diagnóstico , Neoplasias Gástricas/genética , Espectroscopia de Infravermelho com Transformada de Fourier , Técnicas Biossensoriais/métodos , DNA de Cadeia Simples/genética , MicroRNAs/genética , Zinco , Técnicas Eletroquímicas/métodos , Limite de DetecçãoRESUMO
In the present study, a label-free electrochemical genosensor was designed based on ZnS quantum dots functionalized with l-cysteine (Cys-ZnS-QDs) to detect miR-200a, as a special ovarian cancer biomarker. The Cys-ZnS-QD genosensor was characterized by transmission electron microscopy (TEM), UV-Vis absorption and fluorescence methods. Cys-ZnS-QDs are electrodeposited on the glassy carbon electrode surface and act as a suitable substrate for immobilization of the DNA probe. The effective parameters in the preparation of the genosensor are optimized to improve its analytical performance. The analytical performance of the genosensor has been investigated using electrochemical impedance spectroscopy. Under optimal conditions, the linear range and the detection limit of miR-200a were found to be 1.0 × 10-14 to 1.0 × 10-6 M and 8.4 fM. In addition, the genosensor is used to detect the target complementary miRNA strand from a single-base mismatch miRNA strand. Finally, this label-free electrochemical biosensor was used to detect miR-200a in human plasma without using any amplification method.