Use of fluorescence resonance energy transfer for rapid detection of enteroviral infection in vivo.

Abstract

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 (2A(pro)), 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 2A(pro) 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.

• Authors: Yu-Chen Hwang, Wilfred Chen, Marylynn V Yates
• Journal: Applied and environmental microbiology (Appl Environ Microbiol) Vol. 72 Issue 5 Pg. 3710-5 (May 2006) ISSN: 0099-2240 [Print] United States
• PMID: 16672521 (Publication Type: Evaluation Study, Journal Article)

Chemical References

• Bacterial Proteins
• Cyan Fluorescent Protein
• Luminescent Proteins
• Peptides
• Viral Proteins
• yellow fluorescent protein, Bacteria
• Green Fluorescent Proteins
• Cysteine Endopeptidases
• picornain 2A, Picornavirus

Topics

• Amino Acid Sequence
• Animals
• Bacterial Proteins (metabolism)
• Cell Line
• Cysteine Endopeptidases (metabolism)
• Enterovirus (pathogenicity, physiology)
• Enterovirus Infections (diagnosis, virology)
• Fluorescence Resonance Energy Transfer (methods)
• Green Fluorescent Proteins (metabolism)
• Luminescent Proteins (metabolism)
• Microscopy, Fluorescence
• Molecular Sequence Data
• Peptides (chemistry, metabolism)
• Poliovirus (pathogenicity, physiology)
• Time Factors
• Viral Proteins (metabolism)
• Virus Replication