Assay method for Vibrio cholerae and Escherichia coli enterotoxins by automated counting of floating chinese hamster ovary cells in culture medium

J Clin Microbiol. 1978 May;7(5):479-85. doi: 10.1128/jcm.7.5.479-485.1978.

Abstract

As Chinese hamster ovary (CHO) cells on plastic proliferate, many cells float off into the medium instead of piling up after they form a monolayer. Fewer cells were floating in the medium when CHO cells were incubated with cholera toxin at a concentration as low as 10 pg/ml. The toxin increased the adhesiveness of the cells forming confluent monolayers so that the floating cells accumulated on the adherent monolayers. On the basis of this finding, a simple, quantitative assay method for cholera and Escherichia coli enterotoxins was devised by cultivating CHO cells in a Linbro multidish and counting the cells in the medium with a Coulter Counter. The method was sensitive enough to detect toxins in 100- to 200-fold-diluted culture media of toxigenic E. coli strains. Little or no activity was detected by this method in the culture medium of nontoxigenic E. coli.

MeSH terms

  • Animals
  • Bacterial Toxins / analysis*
  • Bacterial Toxins / pharmacology
  • Biological Assay / methods*
  • Cell Adhesion / drug effects
  • Cell Line
  • Cells, Cultured / drug effects*
  • Cholera Toxin / analysis*
  • Cholera Toxin / pharmacology
  • Cricetinae
  • Enterotoxins / analysis*
  • Enterotoxins / pharmacology
  • Escherichia coli / analysis*

Substances

  • Bacterial Toxins
  • Enterotoxins
  • Cholera Toxin