A simple and convenient microtiter plate assay for the detection of bactericidal antibodies to Vibrio cholerae O1 and Vibrio cholerae O139.
It is believed that the correlate of protection for cholera can be determined by the serum vibriocidal assay. The currently available vibriocidal assays, based on the conventional agar plating technique, are labor intensive. We developed a simple and convenient microtiter plate assay for the detection of vibriocidal antibodies that is equally as efficient for Vibrio cholerae O1 and for V. cholerae O139. The addition of succinate and neotetrazolium made it possible to measure the growth of surviving bacterial target cells by monitoring a color change. We evaluated assay parameters (target strains, growth of target cells, complement source and concentration) that may affect the reproducibility of the method for V. cholerae O139. The results obtained with the microtiter plate assay were uniformly similar to those obtained with the conventional agar plating assay, when testing both the Inaba and Ogawa serotypes of V. cholerae O1. The microtiter plate assay was also convenient for measuring the activity of animal sera and mouse monoclonal antibodies.
Boutonnier
Alain
A
Unité du Choléra et des Vibrions, Centre National de Référence des Vibrions et du Choléra, Institut Pasteur, 75724 Paris Cedex 15, France.
Dassy
Bruno
B
Duménil
Rémy
R
Guénolé
Alain
A
Ratsitorahina
Maherisoa
M
Migliani
René
R
Fournier
Jean-Michel
JM
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D015941:Complement Hemolytic Activity Assay / Q000379:methods*; D006801:Humans; D007202:Indicators and Reagents / Q000378:metabolism; D051379:Mice; D011817:Rabbits; D018709:Statistics, Nonparametric; D019802:Succinic Acid / Q000378:metabolism; D013778:Tetrazolium Salts / Q000378:metabolism; D041241:Vibrio cholerae O1 / Q000276:immunology*; D041242:Vibrio cholerae O139 / Q000276:immunology*
D000907:Antibodies, Bacterial; D007202:Indicators and Reagents; D013778:Tetrazolium Salts; C011314:neotetrazolium; D019802:Succinic Acid