Development of a colorimetric RT-LAMP amplification assay adapted to an early and easy detection of Dengue virus

CARRILLO C; WERBAJH S; MALNERO C; STOLOWICS FG; L. LAROCCA; MARILAT V; VOJNOV A

Congreso; 18th International Congress on Infectious Diseases & XVIII Congreso SADI; 2018

Dengue virus (DENV) is the most prevalent arbovirus in more than 100 countries within tropical and subtropical regions of the world. There are four distinct serotypes, DENV1, DENV2, DENV3, and DENV4. DENV infection causes dengue fever, the more dangerous dengue hemorrhagic fever and dengue shock syndrome, all of which are very contagious. Early and rapid detection of DENV infection during the acute phase is crucial for proper patient management and to prevent the infection spreading. Several immunoassay test for DENV diagnosis have been developed, however they present mild specificity because of the presence of cross-reactive antigens shared by flaviviruses. Nucleic acid detection is deemed to be an alternative effective method in early DENV infection stage. Reverse transcriptase (RT)-PCR and real-time PCR methods have both been used widely for laboratory diagnosis because of their high sensitivities and specificities; however, they are restricted for specialized laboratories. However, the need for expensive and complex thermal cyclers in addition to the cold storage requirement for the PCR reagents has significantly hampered the use of these methods in resource-limited countries and Point of Care settings. Hence, the loop-mediated isothermal amplification (LAMP) is potentially ideal to be used especially in areas with limited resources. Loop-mediated isothermal amplification (LAMP) originally developed by Notomi et al. (2000) represents a very sensitive, easy to use, and less time consuming diagnostic method. The simplicity and high efficiency of LAMP to rapidly amplify nucleic acids under isothermal conditions suggested that LAMP could be a potential alternative for detecting dengue virus especially in field settings as compared to qRT-PCR.