Dual electrochemical DNA sensor utilizing Ag(I)-Stabilized DNA mismatches for simultaneous, label-free detection of mpox and HIV

The WHO declared Mpox a public health emergency of international concern due to the resurgence of Mpox clade I outbreaks. Following, the CDC reported a high prevalence of HIV co-infection during the global MPOX outbreak. In 2025, no commercially available multiplex PCR assays enable concurrent detection of mpox and HIV. To address these limitations, this study presents a novel dual electrochemical DNA (eDNA) biosensor for the simultaneous, label-free detection of Mpox and HIV nucleic acids using a mismatched-driven nucleic acid–metal ion nanocomplex (MDN) strategy. The sensor exploits C–C mismatches that selectively coordinate with Ag⁺ ions, enabling covalent target capture and sensitive signal generation. It demonstrates a dynamic range of 10¹–10⁵ pM with picomolar-level LODs and successful detection in clinical saliva and urine samples, with LOD below 4 copies/mL. This multiplex platform offers a rapid, cost-effective, and robust diagnostic tool for co-infection screening in high-risk populations.