Failure to detect cell-associated enterotoxin B in Staphylococcus aureus by immunofluorescence.
Enterotoxin B-producing and -nonproducing Staphylococcus aureus strains showed cell fluorescence when tested with fluoresceinisothiocyanate-labeled rabbit anti-enterotoxin B globulin, probably as a result of a protein A-immunoglobulin G (Ig G) interaction. No cell-bound enterotoxin B could be detected by immunofluorescence using F(ab(1))(2)-fragments of anti-enterotoxin B globulin. However, soluble enterotoxin B could be estimated by immunofluorescence. Approximately 1,000-fold more enterotoxin B was detected by immunodiffusion as an extracellular product in the media than could be detected in the cell fraction. The results show that intact Ig G is not suitable for the detection of antigens other than protein A on the cell surface of S. aureus in conventional immunofluorescence. For such purposes, the use of F(ab(1))(2)-fragments of Ig G is recommended.
D000645:Ammonium Sulfate; D000818:Animals; D000937:Antigen-Antibody Reactions; D000942:Antigens, Bacterial / Q000032:analysis; D001426:Bacterial Proteins / Q000032:analysis; D011232:Chemical Precipitation; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D003470:Culture Media; D004586:Electrophoresis; D004768:Enterotoxins / Q000032:analysis* / Q000302:isolation & purification; D005452:Fluoresceins; D005455:Fluorescent Antibody Technique; D007106:Immune Sera; D005779:Immunodiffusion; D007122:Immunoelectrophoresis; D007074:Immunoglobulin G / Q000302:isolation & purification; D008248:Lysostaphin; D008851:Micropore Filters; D010434:Pepsin A; D011817:Rabbits / Q000276:immunology; D013210:Staphylococcus / Q000032:analysis* / Q000254:growth & development / Q000276:immunology; D013861:Thiocyanates
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D003470:Culture Media; D004768:Enterotoxins; D005452:Fluoresceins; D007106:Immune Sera; D007074:Immunoglobulin G; D013861:Thiocyanates; D010434:Pepsin A; D008248:Lysostaphin; D000645:Ammonium Sulfate