Part I: Non-faradaic electrochemical impedance-based DNA biosensor for detecting phytopathogen - Ralstonia solanacearum

Herein, we report for the first time the development of a label-free, non-faradaic, and highly sensitive DNA-based impedimetric sensor using micro-sized gold interdigitated electrodes (IDE) to detect a soil-borne agricultural pathogen Ralstonia solanacearum. A universal 30 oligomer single-stranded DNA (ssDNA) probe lpxC4 having specificity towards R. solanacearum is successfully immobilized on the surface of IDE along with mercaptohexanol. The electrochemical stability of the developed sensor surface is determined using open circuit potential measurements. The DNA probe immobilization protocol is validated using the changes configured on the surface of IDE by contact angle and ATR-FTIR analysis. The DNA target hybridization is detected using non-faradaic electrochemical impedance spectroscopy measurement with an ultra-low sample volume of 10 µL. The non-faradaic approach is verified by studying redox behavior using cyclic voltammetry. We investigate the hybridization of the surface-immobilized label-free probe with the complementary DNA targets obtained from infected eggplant saplings and cross-reactive studies with mismatched DNA strands. Our impedimetric sensor can detect target concentrations as low as 0.1 ng/µL. This standardization and detection of DNA hybridization serves as part I of the work and paves the way for further study in the detection of pathogenic field samples.