Competitive, enzyme-linked immunosorbent assay for toxic shock syndrome toxin 1.
We developed a competitive, enzyme-linked immunosorbent assay for the quantitation of toxic shock syndrome toxin 1 (TSST-1). Polyvalent immunoglobulin G from immunized rabbits was used as the capture antibody, and alkaline phosphatase conjugated to purified toxin served as the indicator enzyme. A standard curve was generated with each experiment, from which the concentration of toxin in culture supernatants was extrapolated. The assay was useful for determining toxin concentrations of 0.03 to 0.5 micrograms/ml, which is a substantial, practical improvement over immunodiffusion methods. Staphylococcal enterotoxins A through E were not significantly cross-reactive in the assay, and staphylococcal protein A did not interfere with quantitation of TSST-1. By testing a variety of staphylococcal strains, we found 100% concordance between toxin determinations made with our assay and those made by the investigators from whom the strains were obtained. The competitive, enzyme-linked immunosorbent assay is a highly reproducible, inexpensive means of determining TSST-1 concentrations and may have broad applicability in the field of toxic shock research.
D000469:Alkaline Phosphatase; D000818:Animals; D001427:Bacterial Toxins; D003429:Cross Reactions; D004768:Enterotoxins / Q000032:analysis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005779:Immunodiffusion; D011817:Rabbits; D013045:Species Specificity; D013205:Staphylococcal Protein A; D013211:Staphylococcus aureus; D018089:Superantigens
D001427:Bacterial Toxins; D004768:Enterotoxins; D013205:Staphylococcal Protein A; D018089:Superantigens; C030107:enterotoxin F, Staphylococcal; D000469:Alkaline Phosphatase