A new sensitive bioassay for precise quantification of interferon activity as measured via the mitochondrial dehydrogenase function in cells (MTT-method).
A biological method for precise quantification of interferons has been developed. The method is based upon the dehydrogenase system of the intact target cells, which will normally convert an artificial substrate, MTT, into formazan (blue), which, in turn, can be measured spectrophotometrically. This conversion is greatly reduced by cytocidal viruses in a dose-dependent manner. The protection of target cells by interferon against challenge virus is reflected in a diminished reduction in the production of formazan, thus giving a very precise method for quantification of interferon. The lowest level of detection is around 0.10 international units. The intra- and inter-assay variability appear to be below 10%. The assay, which makes no use of expensive ingredients, is performed in 96-well micro-trays and read in an inexpensive ELISA-scanner.
Berg
K
K
Institute of Medical Microbiology, University of Copenhagen, Denmark.
Hansen
M B
MB
Nielsen
S E
SE
D001681:Biological Assay / Q000379:methods*; D002460:Cell Line; D003588:Cytopathogenic Effect, Viral; D007372:Interferons / Q000032:analysis*; D008928:Mitochondria / Q000201:enzymology*; D010088:Oxidoreductases / Q000032:analysis*; D013053:Spectrophotometry; D014777:Virus Diseases / Q000201:enzymology
D007372:Interferons; D010088:Oxidoreductases