A Label-Free Electrochemical Genosensor for the Rapid Detection of Campylobacter jejuni, C. coli, C. lari and C. upsaliensis

Campylobacter spp. is one of the most common pathogens responsible for gastroenteritis in developed countries and is raising public health concerns worldwide. This work optimized a label-free electrochemical genosensor based on screen-printed gold electrodes (SPAuEs) for the rapid detection of Campylobacter jejuni, C. coli, C. lari and C. upsaliensis. SPAuEs were functionalized with a specific thiolated DNA probe and tested with a ferrocyanide solution for signal production. The optimization of the conditions was obtained using DNA extracted from pure cultures of Campylobacter spp. and negative controls such as Escherichia coli, Listeria innocua, Salmonella spp., and Helicobacter pylori. Cyclic voltammetry (CV) and differential pulse voltammetry (DPV) were compared to assess sensitivity and specificity. The relative change in intensity of the ferrocyanide anodic peak (Ipa) was proportional to the value of Campylobacter spp. DNA concentrations in the range of 1 pg/µL to 104 pg/µL. The limit of detection of our optimized system was 1.06 pg/μL. After optimization, the method was applied to chicken meat samples from the market. The proposed electrochemical DNA biosensor was able to detect Campylobacter jejuni, C. coli, C. lari and C. upsaliensis after selective enrichment and DNA isolation within 60 min of DNA extraction, demonstrating its usefulness for routine analyses.