Point-of-care detection of Monkeypox virus clades using high-performance upconversion nanoparticle-based lateral flow assay

There is an urgent need for a point-of-care testing (POCT) method in developing and underserved regions to distinguish between two Monkeypox virus (MPXV) clades, given their varying transmissibility and clinical manifestations. In this paper, we target the specific complement protein gene fragment of two MPXV clades and construct a high-performance upconversion nanoparticles–based lateral flow assay (UCNPs-based LFA) with double T-lines and a shared C-line. This enables qualitative and quantitative dual-mode detection when combined with a smartphone and a benchtop fluorescence analyzer. The developed LFA exhibits stable performance, convenient operation, rapid readout (within 8 min), and a much lower limit of detection (LOD) (~ pM level) compared to existing POCT methods. The proposed detection platform demonstrates significant potential for pathogen diagnosis using a POCT approach.