Caister Academic Press

Real-Time NASBA

Authors: Sam Hibbitts and Julie D. Fox
Abstract: NASBA is an isothermal nucleic acid amplification method that is particularly suited to detection and quantification of genomic, ribosomal or messenger RNA. The product of NASBA is single-stranded RNA of opposite sense to the original target. The first developed NASBA methods relied on liquid or gel-based probe-hybridisation for post-amplification detection of products. More recently, real-time procedures incorporating amplification and detection in a single step have been reported and applied to a wide range of targets. Thus real-time NASBA has proved to be the basis of sensitive and specific assays for detection, quantification and analysis of RNA (and in one case DNA) targets. Molecular beacons have been utilised for detection of NASBA products in all published real-time procedures whether for commercially-available kits or for in-house diagnostic assays. As experience in design of such fluorescent-labelled probes increases and fluorimeters suitable for their detection become widely available, real- time NASBA methodology will be confirmed as a suitable alternative to other real-time amplification methods such as reverse transcriptase PCR (RT-PCR).
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