Detection of human TNF-alpha mRNA by NASBA.
A method to amplify and detect TNF-alpha mRNA from primed Mono Mac 6 cells is described. A silica-based extraction system was utilised for preparation of cell extracts and specific oligonucleotide primers were designed for amplification of TNF-alpha mRNA by the NASBA process. Amplification products were detected using either a liquid hybridisation assay, with analysis by polyacrylamide gel electrophoresis, or a plate hybridisation system. The method has many potential applications for the study of inflammatory cytokines and cellular mRNAs in cell culture and clinical samples.
Darke
B M
BM
Department of Medical Microbiology, University of Wales College of Medicine, Heath Park, Cardiff, UK.
Jackson
S K
SK
Hanna
S M
SM
Fox
J D
JD
D002478:Cells, Cultured; D017931:DNA Primers; D006801:Humans; D008264:Macrophages / Q000166:cytology; D009000:Monocytes / Q000166:cytology; D021141:Nucleic Acid Amplification Techniques; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000032:analysis*; D012680:Sensitivity and Specificity; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000235:genetics*
D017931:DNA Primers; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha