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Flow cytometry-based assay for titrating dengue virus.

C R Lambeth¹, L J White, R E Johnston

¹Department of Microbiology and Immunology, CB #7290, University of North Carolina, Chapel Hill, NC 27599, USA.

Journal of Clinical Microbiology

|July 8, 2005

Summary

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A new fluorescence-activated cell sorting (FACS) assay offers a faster method for quantifying dengue virus (DENV) infections. This FACS assay provides accurate dengue virus titration comparable to traditional methods but in significantly less time.

Area of Science:

Virology
Immunology
Biotechnology

Background:

Traditional plaque assays for dengue virus (DENV) titration are laborious and may fail with certain clinical isolates.
Fluorescence-activated cell sorting (FACS) is a sensitive method for detecting DENV-infected cells.

Purpose of the Study:

To develop and validate a FACS-based assay for efficient and accurate dengue virus titration.
To compare the performance of the FACS assay with conventional plaque and endpoint dilution assays.

Main Methods:

DENV-infected cells were quantified using FACS at 24 hours post-infection.
FACS-derived titers were compared against plaque and endpoint dilution assay results for various DENV strains.
The assay's ability to titrate clinical DENV isolates was assessed.

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Main Results:

The number of infected cells detected by FACS at 24 hours post-infection correlated with the first round of infection, serving as a reliable DENV titer readout.
FACS assay titers were comparable to plaque and endpoint dilution assay titers for most laboratory and clinical DENV strains.
The FACS assay successfully titrated clinical DENV isolates that do not form plaques.

Conclusions:

The developed FACS assay provides a significantly faster (24 hours vs. 5–7 days) and more versatile alternative to plaque assays for DENV titration.
This method is suitable for a broader range of DENV strains, including challenging clinical isolates.
The FACS assay streamlines dengue virus research by improving the efficiency of viral quantification.