Adaptation of a colorimetric microtitration assay for quantifying Pasteurella haemolytica A1 leukotoxin and antileukotoxin

A colorimetric microtitration assay was adapted to quantify the cytotoxicity of Pasteurella haemolytica A1 leukotoxin to bovine neutrophils used as target cells. The viability of leukotoxin-treated target cells was detected by use of a tetrazolium dye that living cells reduced to dark blue formazan....

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Bibliographic Details
Published inAmerican journal of veterinary research Vol. 48; no. 11; p. 1559
Main Authors Vega, M.V, Maheswaran, S.K, Leininger, J.R, Ames, T.R
Format Journal Article
LanguageEnglish
Published United States 01.11.1987
Subjects
Animals
ANTITOXINS
BOVIN
CATTLE
Cattle - blood
COLORIMETRIA
COLORIMETRIE
COLORIMETRY
Colorimetry - methods
CYTOTOXICITY
Exotoxins - analysis
Exotoxins - antagonists & inhibitors
Exotoxins - pharmacology
GANADO BOVINO
LEUCOCITOS
LEUCOCYTE
LEUKOCYTES
Neutrophils - drug effects
PASTEURELLA
TOXICIDAD
TOXICITE
TOXICITY
TOXINAS
TOXINE
TOXINS
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Summary:A colorimetric microtitration assay was adapted to quantify the cytotoxicity of Pasteurella haemolytica A1 leukotoxin to bovine neutrophils used as target cells. The viability of leukotoxin-treated target cells was detected by use of a tetrazolium dye that living cells reduced to dark blue formazan. The amount of formazan formed (which was quantified by use of an ELISA plate reader) was directly proportional to the number of viable target cells. This assay system also was used to measure leukotoxin-neutralization antibody titers of bovine serum and lung lavage specimens obtained during vaccination experiments. The major advantages of this assay over other methods such as the 51Cr-release and trypan blue-exclusion assays are precision, rapidity, and low cost; it also does not use radioisotopes.
Bibliography:L73
881114488
ISSN:0002-9645
1943-5681