(598a) Molecular Beacons for Parallel Diagnosis of Infectious Viruses

The emergence new viral agents in conjunction with the traditional agents such as influenza necessitate their rapid identification for the best line of treatment. Since the conventional culture, immuno staining, and PCR methods are time consuming and expensive, a new method is needed that can provide automated identification of multiple viral agents. Among the technologies currently under development for real-time detection, the most promising one is molecular beacons (MBs). We have recently reported the use of nuclease-resistant MBs containing 2'-O-methyl RNA bases with phosphorothioate internucleotide linkages for the real-time detection of coxsackievirus B6 (CVB6) replication in living cells via TAT peptide delivery. Presence of the TAT peptide enabled nearly 100% intracellular delivery within 15 min and a discernible fluorescence signal was observed at 30 min post-infection. These results suggest that the TAT-modified, nuclease-resistant MBs when coupled with flow cytometry could be a powerful tool for real-time detection of infected cells within 15 to 30 min. In this talk, recently progress in this area will be presented using poliovirus infection as a model.