Detection of Dengue Virus Serotype 3 Using a Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification Assay: Evaluation with Clinical Samples from Southeastern Mexico

Dengue virus (DENV), an important mosquito-borne orthoflavivirus, represents a growing global threat due to its geographic expansion and recent outbreaks worldwide. In resource-limited endemic settings, the development of affordable diagnostic assays is needed. In this study, we developed and validated a colorimetric reverse transcription loop-mediated isothermal amplification assay (RT-LAMP) for the detection of DENV type 3 (DENV-3) using 95 previously diagnosed clinical samples from Southeastern Mexico. Primers targeting the 3′ untranslated region (3′ UTR) of DENV-3 were designed, and assay conditions were standardized. The colorimetric RT-LAMP DENV-3 system achieved a preliminary limit of detection of 1 × 103 copies per reaction, with 90.7% sensitivity and 100% specificity. The colorimetric format enabled visual readout without specialized equipment, supporting its potential applicability in point-of-care and resource-limited settings. The developed colorimetric RT-LAMP detection for DENV-3 is intended as a rapid screening/triage tool that can trigger confirmatory testing or public-health actions.