The outbreak of the monkeypox epidemic underscores the importance of developing a rapid and sensitive virus detection technique. Microneedles (MNs) offer minimally invasive sampling capabilities, providing a solution for the development of integrated extraction and diagnostic portable devices. Here, we report an integrated MNs and hydrogel biosensor (IMHB) platform, composed of an electronic device, an MN patch, and a hydrogel patch. The IMHB allowed for specific extraction of monkeypox virus (MPXV) directly from lesional skin and virus detection in both electrochemical and colorimetric modes. A bifunctional signal probe 3,3',5,5'-tetramethylbenzidine (TMB) was loaded in a hydrogel patch, providing measurable signals for dual-mode sensing. Additionally, a control area was designed in this platform to collect blank samples from normal skin, enabling ratio analysis and quality control functions. This dual-mode ratiometric sensing strategy exhibited a wide range of 10-1000 ng/mL for MPXV A29 protein, with detection limits of 0.1632 and 0.3017 ng/mL for electrochemical and colorimetric assay, respectively. The developed IMHB platform provides a novel way for rapid on-site determination of MPXV, demonstrating the potential for quick intervention in the early stages of infectious diseases.
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