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Journal of Clinical Microbiology, July 2005, p. 3267-3272, Vol. 43, No. 7
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.7.3267-3272.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Flow Cytometry-Based Assay for Titrating Dengue Virus

Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, North Carolina

Received 3 September 2004/ Returned for modification 16 January 2005/ Accepted 3 March 2005

Plaque assays for titrating dengue virus (DENV) are time-consuming and not suitable for strains that do not plaque. Fluorescence-activated cell sorting (FACS) has been used to detect DENV-infected cells. Here we describe a FACS-based assay for titrating DENV. We determined that at 24 h postinfection, the number of infected cells detected by FACS represented the first round of infection and therefore could be used as a readout of the number of infectious particles in the inoculum. When the titers of different laboratory and clinical strains of DENV were compared using FACS, plaque, and endpoint dilution assays, for most strains the FACS titers were comparable to titers obtained by plaque or endpoint dilution assays. The FACS assay is an improvement over the plaque assay because the infection period is reduced from 5 to 7 days to 24 h and the assay can be used to titrate clinical isolates that frequently do not form clear plaques on cell monolayers. The novel FACS-based methods described here will facilitate laboratory studies of dengue.

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