A colorimetric microassay for opsonins by reduction of NBT in phagocytosing bovine polymorphs

J Immunol Methods. 1986 Jun 24;90(2):197-201. doi: 10.1016/0022-1759(86)90076-1.

Abstract

The adaptation of the reduction of nitro blue tetrazolium (NBT) by phagocytes to the measurement of the opsonising capacity of serum or milk whey is described. A dose-response curve of the absorbance of solubilised reduced NBT to the concentration of opsonins, along with a close correlation (r = 0.94) between the colorimetric NBT assay and ingestion of group B streptococci (as measured by reduction of [3H]thymidine incorporation by extracellular bacteria), was established. This NBT reduction test, employing 96-well flat-bottom microtitre plates, is simple, semiautomated, requires a low number of PMN, and is applicable to a large number of samples, which renders it useful as a screening test for the opsonising capacity of sera or other biologic fluids.

MeSH terms

  • Animals
  • Cattle
  • Neutrophils / physiology*
  • Nitroblue Tetrazolium*
  • Opsonin Proteins / analysis*
  • Oxidation-Reduction
  • Phagocytosis*
  • Streptococcus agalactiae / immunology
  • Tetrazolium Salts*

Substances

  • Opsonin Proteins
  • Tetrazolium Salts
  • Nitroblue Tetrazolium