MOLECULAR BEACON PROBES COMBINED WITH AMPLIFICATION BY NASBA ENABLE HOMOGENEOUS, REAL-TIME DETECTION OF RNA

Molecular beacon probes can be employed in a NASBA amplicon detection system to generate a specific fluorescent signal concomitantly with am plification. A molecular beacon, designed to hybridize within the targ et sequence, was introduced into NASBA reactions that amplify the geno mic RNA of potato leaf roll virus (PLRV). During amplification, the pr obe anneals to the antisense RNA amplicon generated by NASBA, producin g a specific fluorescent signal that can be monitored in real-time. Th e assay is rapid, sensitive and specific. As RNA amplification and det ection can be carried out in unopened vessels, it minimizes the risk o f carry-over contaminations. Robustness has been verified on real-worl d samples. This homogeneous assay, called AmpliDet RNA, is a significa nt improvement over current detection methods for NASBA amplicons and is suitable for one-tube applications ranging from high-throughput dia gnostics to in vivo studies of biological activities.