Dengue Virus: Isolation, Propagation, Quantification, and Storage

Abstract 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 manifestations of dengue can have a wide range of outcomes varying from a mild febrile illness to a life‐threatening condition. New techniques have largely replaced the use of DENV isolation in disease diagnosis. However, virus isolation still serves as the gold standard for detection and serotyping of DENV and is common practice in research and reference laboratories where clinical isolates of the virus are characterized and sequenced, or used for a variety of research experiments. Isolation of DENV from clinical samples can be achieved in mammalian and mosquito cells or by inoculation of mosquitoes. The experimental methods presented here describe the most common procedures used for the isolation, serotyping, propagation, and quantification of DENV. Curr. Protoc. Microbiol . 27:15D.2.1‐15D.2.24. © 2012 by John Wiley & Sons, Inc.