Development of a Multi-Target TaqMan Assay to Detect Eastern Equine Encephalitis Virus Variants in Mosquitoes

作者：Armstrong Philip M^*; Prince Nicholanna; Andreadis Theodore G

来源：Vector Borne and Zoonotic Diseases, 2012, 12(10): 872-876.

DOI：10.1089/vbz.2012.1008

• Eastern equine encephalitis virus
• Real-time PCR
• Surveillance
• TaqMan

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.

• 出版日期2012-10