A novel quantitative multiplex NASBA method: application to measuring tissue factor and CD14 mRNA levels in human monocytes

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...

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Veröffentlicht in:	Nucleic acids research 1999-09, Vol.27 (17), p.e15-15
Hauptverfasser:	van Deursen, P B, Gunther, A W, van Riel, C C, van der Eijnden, M M, Vos, H L, van Gemen, B, van Strijp, D A, Tackent, N M, Bertina, R M
Format:	Artikel
Sprache:	eng
Schlagworte:	Biological Assay - methods Cells, Cultured Genetic Markers Humans Lipopolysaccharide Receptors - genetics Lipopolysaccharides - immunology Monocytes - immunology Monocytes - metabolism Nucleic Acid Amplification Techniques Reproducibility of Results RNA, Messenger - analysis Thromboplastin - genetics Time Factors
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Zusammenfassung:	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.
ISSN:	1362-4962 0305-1048 1362-4962
DOI:	10.1093/nar/27.17.e15