Journal
VECTOR-BORNE AND ZOONOTIC DISEASES
Volume 12, Issue 10, Pages 872-876Publisher
MARY ANN LIEBERT, INC
DOI: 10.1089/vbz.2012.1008
Keywords
Eastern equine encephalitis virus; Real-time PCR; Surveillance; TaqMan
Funding
- Centers for Disease Control and Prevention [U50/CCU116806-01-1]
- U.S. Department of Agriculture [CONH00768, CONH00773]
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Disease outbreaks caused by eastern equine encephalitis virus (EEEV; Togaviridae, Alphavirus) may be prevented by implementing effective surveillance and intervention strategies directed against the mosquito vector. Methods for EEEV detection in mosquitoes include a real-time reverse transcriptase PCR technique (TaqMan assay), but we report its failure to detect variants isolated in Connecticut in 2011, due to a single base-pair mismatch in the probe-binding site. To improve the molecular detection of EEEV, we developed a multi-target TaqMan assay by adding a second primer/probe set to provide redundant targets for EEEV detection. The multi-target TaqMan assay had similar performance characteristics to the conventional assay, but also detected newly-evolving strains of EEEV. The approach described here increases the reliability of the TaqMan assay by creating back-up targets for virus detection without sacrificing sensitivity or specificity.
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