Light-Switch Electrochemiluminescence-Driven microfluidic sensor for rapid and sensitive detection of Mpox virus

Abstract

Rapid, real-time, and accurate detection of pathogenic nucleic acids is crucial for the control and treatment of Mpox virus infection; however, classical detection methods are limited by time-consuming amplification steps, cumbersome equipment, and the requirement for professional operators. Herein, we developed an electrochemiluminescence (ECL)-driven microfluidic detection chip based on a light-switch molecule, which when coupled with rapid recombinase polymerase amplification (RPA) enabled the detection of Mpox within 30 min. [Ru(phen)2dppz]2+ was used as the ECL light-switch molecule owing to its significant ECL signal enhancement property when intercalated into double-stranded RPA products. The microfluidic chip included two independent liquid storage tanks preloaded with an RPA reagent mixture and [Ru(phen)2dppz]2+, which were mixed via centrifugation to produce ECL signals. The developed sensing platform effectively detected Mpox virus with high sensitivity, and the limit of detection was as low as 10 copies/µL. Moreover, it showed 100 % sensitivity and 100 % specificity when validated against the quantitative polymerase chain reaction assay using clinical samples from the pustules, throat, urine, blood, saliva, and anus of Mpox-positive patients. Combined with a hand-held detection device, this platform can facilitate the early diagnosis of Mpox cases in point-of-care testing without trained personnel and specialized equipment.