Development of a Simple and Rapid Assay for Field Detection of Dengue Viral RNA

Dengue virus is a mosquito born flavivirus that represents a major threat to military forces deployed to tropical areas of the world. Accurate and efficient diagnosis of dengue is important for military health, clinical care, epidemiology, pathogenesis studies, and vaccine research. Currently, there are several PCR-based methods for detecting viral nucleic acid in serum, and this technique is frequently used in the case of the Dengue virus. However, the loop-mediated isothermal amplification (LAMP) technique provides a novel, easy-to-use alternative. Unlike conventional PCR which requires a thermocycler, LAMP reactions are held at a constant temperature and the read out can be done with the naked eye. There are sufficient common regions in the genomes of all four strains to develop ‘pan-dengue reagents”. In this project, a generic LAMP protocol will be established, and investigations into the development of LAMP probes will be done to allow the specific identification of each dengue strain.