Dual targets-induced specific hemin/G-quadruplex assemblies for label-free electrochemical detection capable of distinguishing Salmonella and its common serotype in food samples

Abstract

Efficient detection of pathogenic bacteria is paramount for ensuring food safety and safeguarding public health. Herein, we developed a label-free and signal-on dual-target recognition electrochemical DNA sensing platform based on the conformational formation of split G-quadruplex. This platform focused on achieving sensitive and low-cost detection of Salmonella and its most human-infecting S. typhimurium serotype. In simple terms, the dual-target recognition probe (DTR-6P) was ingeniously designed for the loop sequence on the loop-mediated isothermal amplification (LAMP) amplicons. It could recognize two different genes and release their corresponding G-rich sequences. The exfoliated G-rich sequences could be captured by the capture probes on the electrode, and then the bimolecular G-quadruplex or the tetramolecular G-quadruplex would be formed to capture hemin, thereby enabling dual-signal reporting. The minimum detection amount of target genes can be as low as 2 copies/μL. Encouragingly, the real food samples contaminated by Salmonella and the S. typhimurium serotype can be readily identified. The sensing platform with ingenious design paves a new way for label-free, multi-target simultaneous detection, whose advantage of rapidity, sensitivity, cost-effectiveness, and specificity also lay a solid foundation for practical applications.

Keywords: Electrochemical biosensor; Label-free; Multiplex detection; Salmonella; Split G-quadruplex.