Development and Validation of a Combined RT-LAMP Assay for the Rapid and Sensitive Detection of Dengue Virus in Clinical Samples from Colombia

Background: Dengue virus (DENV) infection is a significant public health concern in many trop-ical and subtropical regions, where early and rapid detection is crucial for effective patient man-agement and controlling the spread of the disease. Particularly in resource-limited, rural healthcare settings where dengue is endemic, there is a need for diagnostic methods that are both easy to perform and highly sensitive. Objective: This study focuses on the development and validation of a single-tube reverse tran-scription-loop-mediated isothermal amplification (RT-LAMP) assay for the detection of DENV. Methodology: The RT-LAMP assay designed in this work combines two sets of primers targeting the 5' and 3'-UTR regions of the dengue virus, aiming to increase the sensitivity of detection. Results: The clinical validation of this COMB-RT-LAMP in samples from febrile individuals with a serological or antigenic diagnosis showed a sensitivity of over 95%. The assay's perfor-mance was statistically compared to standard diagnostic method quantitative reverse transcrip-tion-polymerase chain reaction (qRT-PCR). Conclusions: The findings support the potential of RT-LAMP as a rapid, sensitive, and specific tool for dengue diagnosis and surveillance, particularly suitable for field use in low-resource set-tings.