title: A novel quantitative multiplex NASBA method: application to measuring tissue factor and CD14 mRNA levels in human monocytes.
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.
authors.affiliation: Organon Teknika B.V., Boseind 15, PO Box 84, 5280 AB Boxtel, The Netherlands. p.deursen@teknika.btl.akzonobel.nl
authors.forename: P B
authors.initials: PB
authors.lastname: van Deursen
authors.forename: A W
authors.initials: AW
authors.lastname: Gunther
authors.forename: C C
authors.initials: CC
authors.lastname: van Riel
authors.forename: M M
authors.initials: MM
authors.lastname: van der Eijnden
authors.forename: H L
authors.initials: HL
authors.lastname: Vos
authors.forename: B
authors.initials: B
authors.lastname: van Gemen
authors.forename: D A
authors.initials: DA
authors.lastname: van Strijp
authors.forename: N M
authors.initials: NM
authors.lastname: Tackent
authors.forename: R M
authors.initials: RM
authors.lastname: Bertina
mesh_terms: D001681:Biological Assay / Q000379:methods; D002478:Cells, Cultured; D005819:Genetic Markers; D006801:Humans; D018950:Lipopolysaccharide Receptors / Q000235:genetics*; D008070:Lipopolysaccharides / Q000276:immunology; D009000:Monocytes / Q000276:immunology / Q000378:metabolism*; D021141:Nucleic Acid Amplification Techniques; D012333:RNA, Messenger / Q000032:analysis*; D015203:Reproducibility of Results; D013925:Thromboplastin / Q000235:genetics*; D013997:Time Factors
chemical_list: D005819:Genetic Markers; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D013925:Thromboplastin