Real-Time NASBA
Julie D. Fox, Catherine Moore and Diana Westmoreland
from: Real-Time PCR: Current Technology and Applications (Edited by: Julie Logan, Kirstin Edwards and Nick Saunders). Caister Academic Press, U.K. (2009)
Abstract
NASBA is an isothermal nucleic acid amplification method which 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. 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 RNA and some DNA targets. Thus real-time NASBA has proved to be the basis of sensitive and specific assays for detection, quantification and differentiation of RNA and DNA targets. Molecular beacons have most often been utilised in real-time NASBA whether in commercially-available kits or as published in-house developed assays. As experience in design of molecular-beacon probes increases and fluorimeters suitable for real-time NASBA become widely available this methodology will be confirmed as a suitable alternative to real-time RT-PCR (and perhaps DNA PCR) read more ...
