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Applied and Environmental Microbiology, May 2006, p. 3710-3715, Vol. 72, No. 5
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.5.3710-3715.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Use of Fluorescence Resonance Energy Transfer for Rapid Detection of Enteroviral Infection In Vivo{dagger}

Yu-Chen Hwang,1 Wilfred Chen,2 and Marylynn V. Yates1*

Department of Environmental Sciences,1 Department of Chemical and Environmental Engineering, University of California, Riverside, California2

Received 28 October 2005/ Accepted 2 March 2006

Enteroviruses can be easily transmitted through the fecal-oral route and cause a diverse array of clinical manifestations. Recent outbreaks associated with enteroviral contamination in aquatic environments have called for the development of a more efficient and accurate virus monitoring system. To develop a simple, rapid, and direct method for identifying enteroviral infections, we generated a fluorescent reporter system in which genetically engineered cells express a hybrid fluorescent indicator composed of a linker peptide, which is exclusively cleaved by the 2A protease (2Apro), flanked with a cyan fluorescent protein (CFP) and a yellow fluorescent protein undergoing fluorescence resonance energy transfer. The covalent linkage between two fluorophores is disrupted due to 2Apro activity upon viral infection, which results in an increase in CFP intensity. This allows the rapid (within 7.5 h) detection of very low numbers (10 PFU or fewer) of infectious enteroviruses.

* Corresponding author. Mailing address: Department of Environmental Sciences, University of California, Riverside, CA 92521. Phone: (951) 827-2358. Fax: (951) 827-3993. E-mail: marylynn.yates{at}ucr.edu.

{dagger} Supplemental material for this article may be found at http://aem.asm.org.

Applied and Environmental Microbiology, May 2006, p. 3710-3715, Vol. 72, No. 5
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.5.3710-3715.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.





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Copyright © 2006 by the American Society for Microbiology. All rights reserved.