A novel quantitative multiplex NASBA method: application to measuring tissue factor and CD14 mRNA levels in human monocytes
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Characterization of Human Monocyte Subsets by Whole Blood Flow Cytometry Analysis
Published on: October 17, 2018
P B van Deursen1, A W Gunther, C C van Riel
1Organon Teknika B.V., Boseind 15, PO Box 84, 5280 AB Boxtel, The Netherlands. p.deursen@teknika.btl.akzonobel.nl
Abstract:
A new method to quantify two individual mRNAs in a single NASBA reaction is described. In this study, tissue factor and CD14 mRNAs were used as a model system. RNA ratios of -4 to +4 log units were determined with good precision (within 0.3 log) and accuracy (within 0.2 log). By measuring both mRNAs in human monocytes that were stimulated with LPS, the multiplex Q-NASBA proved to be a successful tool to monitor the expression levels of two individual mRNAs in a single-tube amplification system. The method has potential in all fields in which quantitative information is needed on two individual RNAs.
