TNF as biomarker for rapid quantification of active Staphylococcus enterotoxin A in food.
Staphylococcus aureus is a major bacterial pathogen which causes clinical infections and food poisoning. This bacterium produces a group of twenty-one enterotoxins (SEs). These enterotoxins have two separate but related biological activities. They cause gastroenteritis and function as superantigens that activate large numbers of T cells. The current method for detection of enterotoxins activity is an in vivo monkey or kitten bioassay; however, this method is not practical to test on a large number of samples. Several immunological assays have been developed however, but these assays cannot distinguish between active toxin which causes food poisoning and inactive toxin, which can bind antibody, but shows no toxicity. The current study demonstrates that short term ex vivo exposure of primary naïve CD4(+) T-cells or splenocytes to SEA induces differential expression and secretion of tumor necrosis factor (TNF) protein. We used immunomagnetic beads coated with anti-SEA antibody to specifically isolate SEA from food. After the eluted toxin was added to the cells SEA biological activity was measured by quantifying TNF protein expression or secretion.
Western Regional Research Center, Agricultural Research Service, US Department of Agriculture, Albany, CA 94710, USA. reuven.rasooly@ars.usda.gov
Reuven
R
Rasooly
Bradley
B
Hernlem
D000818:Animals; D015415:Biomarkers / Q000032:analysis*; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects; D004768:Enterotoxins / Q000032:analysis* / Q000633:toxicity; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005506:Food Contamination / Q000032:analysis*; D066298:In Vitro Techniques; D051379:Mice; D008810:Mice, Inbred C57BL; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis*
D015415:Biomarkers; D004768:Enterotoxins; D014409:Tumor Necrosis Factor-alpha; C026066:enterotoxin A, Staphylococcal