Open AccessEvaluation of a new cytotoxicity assay for Clostridium perfringens type D epsilon toxin
Author(s) -
Payne Dean W.,
Williamson E.Diane,
Havard Helen,
Modi Naren,
Brown Jill
Publication year - 1994
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1994.tb06695.x
Subject(s) - acridine orange , cytotoxicity , clostridium perfringens , toxin , ethidium bromide , formazan , microbiology and biotechnology , viability assay , biology , chemistry , biochemistry , in vitro , bacteria , apoptosis , dna , genetics
A new cytotoxicity assay for determining the activity of epsilon toxin produced by Clostridium perfringens type D has been developed. Viability of cultured cells was determined by the ability of only live cells to convert 5‐(3‐carboxymethoxyphenyl)‐2‐(4,5‐dimethylthiazolyl)‐3‐(4‐sulfophenyl)tetrazolium to the coloured product formazan in the presence of phenazine methosulfate. Of the 12 cell lines tested, only the MDCK cell line was susceptible to epsilon toxin. Specificity was confirmed by the ability of only specific monoclonal antibodies to inhibit cytotoxicity. Good correlation was obtained with the mouse lethality assay ( r = 0.991) and over a wide range of viability (15–75%) as determined by ethidium bromide/acridine orange staining ( r = 0.995).