Validation and application of a visual LAMP assay for Mpox diagnosis — Insights from clinical samples in Thailand

Abstract

Despite numerous recent advances in infectious disease diagnostic technologies including quantitative (q) polymerase chain reaction (PCR) and antigen test kit (ATK), there remains a need to develop more efficient point-of-care assays for large-scale operations. In an effort towards such a screening test for low-resource settings, we have established a colorimetric loop-mediated isothermal amplification assay and detected the amplified DNA via naked-eye observation to maximize assay simplicity using monkeypox (Mpox) as a model. The assay is cheap, highly sensitive and specific to Mpox with the potential for translation into broader applications. • Our protocol offers a simpler, faster, and cheaper test for Mpox than the current gold-standard qPCR, with 100% accuracy. • assay enables decentralized screening and provides visually interpretable results immediately after incubation • The versatility of this protocol allows for easy adaptation to detecting new or re-emerging diseases, utilizing a visual readout for maximum simplicity. © 2025