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 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.
van Deursen
P B
PB
Organon Teknika B.V., Boseind 15, PO Box 84, 5280 AB Boxtel, The Netherlands. p.deursen@teknika.btl.akzonobel.nl
Gunther
A W
AW
van Riel
C C
CC
van der Eijnden
M M
MM
Vos
H L
HL
van Gemen
B
B
van Strijp
D A
DA
Tackent
N M
NM
Bertina
R M
RM
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
D005819:Genetic Markers; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D013925:Thromboplastin