Parsonnet J, Mills J T, Gillis Z A, Pier G B
J Clin Microbiol. 1985 Jul;22(1):26-31. doi: 10.1128/jcm.22.1.26-31.1985.
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.
我们开发了一种用于定量毒性休克综合征毒素1（TSST-1）的竞争性酶联免疫吸附测定法。用免疫兔的多价免疫球蛋白G作为捕获抗体，用与纯化毒素偶联的碱性磷酸酶作为指示酶。每次实验都绘制标准曲线，据此推断培养上清液中毒素的浓度。该测定法可用于测定浓度为0.03至0.5微克/毫升的毒素，与免疫扩散法相比有显著的实际改进。在该测定法中，葡萄球菌肠毒素A至E没有明显的交叉反应，葡萄球菌蛋白A也不干扰TSST-1的定量。通过检测多种葡萄球菌菌株，我们发现用我们的测定法测定的毒素结果与提供菌株的研究者所测定的结果100%一致。这种竞争性酶联免疫吸附测定法是一种高度可重复、低成本的测定TSST-1浓度的方法，可能在毒性休克研究领域有广泛的应用。
