Detection of human TNF-alpha mRNA by NASBA

J Immunol Methods. 1998 Mar 1;212(1):19-28. doi: 10.1016/s0022-1759(97)00211-1.

Abstract

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.

MeSH terms

  • Cells, Cultured
  • DNA Primers
  • Humans
  • Macrophages / cytology
  • Monocytes / cytology
  • Nucleic Acid Amplification Techniques*
  • Nucleic Acid Hybridization
  • RNA, Messenger / analysis*
  • Sensitivity and Specificity
  • Tumor Necrosis Factor-alpha / biosynthesis
  • Tumor Necrosis Factor-alpha / genetics*

Substances

  • DNA Primers
  • RNA, Messenger
  • Tumor Necrosis Factor-alpha