Dengue is a disease caused by infection with one of the four dengue virus serotypes(DENV-1, -2, -3, and -4). The virus is transmitted to humans by Aedes sp. mosquitoes.This enveloped virus contains a positive single-stranded RNA genome. Clinical manifestationsof dengue can have a wide range of outcomes varying from a mild febrile illnessto a life-threatening condition. New techniques have largely replaced the use of DENVisolation in disease diagnosis. However, virus isolation still serves as the gold standardfor detection and serotyping of DENV and is common practice in research and referencelaboratories where clinical isolates of the virus are characterized and sequenced, or usedfor a variety of research experiments. Isolation of DENV from clinical samples can beachieved in mammalian and mosquito cells or by inoculation of mosquitoes. The experimentalmethods presented here describe the most common procedures used for theisolation, serotyping, propagation, and quantification of DENV. Curr. Protoc. Microbiol.27:15D.2.1-15D.2.24