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Am J Trop Med Hyg. 2013 Nov;89(5):1001-5. doi: 10.4269/ajtmh.13-0100. Epub 2013 Sep 9.

Comparison of the mosquito inoculation technique and quantitative real time polymerase chain reaction to measure dengue virus concentration.

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1
Signature Research Program in Emerging Infectious Diseases, Duke-National University of Singapore Graduate Medical School, Singapore, Republic of Singapore.

Abstract

An accurate measure of infectious dengue virus in human and mosquito tissues is critical to fully understand virus-host relationships, disease severity, viral fitness, and pathogenesis. In recent years, RNA copy number measured by quantitative real time-polymerase chain reaction has been used to measure dengue virus concentration in vitro and in vivo. In this study, we detail important differences in the measurement of viral growth kinetics in Vero and C6/36 tissue cultures, in Aedes aegypti mosquitoes, and in viremic human sera using RNA genomic equivalents and mosquito infectious dose 50 (MID50). Although there was reasonably good correlation between the two methods, RNA copy number was 2 to 5 logs greater than infectious virus titers. These differences varied significantly depending on virus strain, viral platform, infectious virus assay, and viral growth phase. The results have important implications for the correct interpretation of biological and epidemiological data from experimental and clinical studies, and show that genomic equivalents should be interpreted with caution when used as a proxy for infectious virus in such studies.

PMID:
24019432
PMCID:
PMC3820311
DOI:
10.4269/ajtmh.13-0100
[Indexed for MEDLINE]
Free PMC Article
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