Qualitative and quantitative methods for detecting staphylococcal epidermolytic toxin.
Qualitative and quantitative methods for detecting staphylococcal epidermolytic toxin.
复制标题
检测葡萄球菌表皮松解毒素的定性和定量方法。
DOI：
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发表时间：
1976
影响因子：
3
通讯作者：
B. Billcliffe
中科院分区：
文献类型：
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作者：
J. Arbuthnott;B. Billcliffe
通过以下方式筛选 98 株金黄色葡萄球菌培养物的浓缩上清液中表皮松解毒素的产生：(1) 3 日龄小鼠的生物学测试，(2) 针对特定抗血清的双扩散沉淀测试，以及 (3) 薄层凝胶等电聚焦上特征蛋白带的出现。在所有这三项测试中，11 种培养物的上清液均获得了阳性结果；当新生小鼠受到活生物体攻击时，这 11 种菌株（没有其他菌株）产生了表皮分裂。在调查中包含的 14 种 II 型噬菌体菌株中，有 8 种（58%）产生表皮松解毒素。三个产毒菌株属于II组以外的噬菌体组。使用针对纯化的表皮松解毒素的抗血清的径向免疫扩散测试证明对于测量超过200微克/毫升的表皮松解毒素的量是令人满意的。免疫扩散测试的结果表明，11 种阳性菌株中的 6 种产生两种血清学上不同形式的表皮松解毒素，其余菌株仅产生其中一种。在薄层等电聚焦凝胶中，血清型 i 和 ii 毒素的存在与蛋白质条带 i 和 ii 的出现之间观察到显着的相关性。这些测试应有助于在未来对从里特型和脓疱病中毒性表皮坏死松解症中分离出的菌株进行调查时，对金黄色葡萄球菌产生不同血清型表皮松解毒素的情况进行定性和定量评估。先前指定为 ETA (pI=7-0) 和 ETB (pI=6-0) 的两种形式的表皮松解毒素是通过 i 型血清毒素的制备型等电聚焦来检测的。有证据表明，热效应的研究应该提供一种研究表皮松解毒素不同分子形式之间关系的方法。
Concentrated supernates of cultures of 98 strains of Staphylococcus aureus were screened for the production of epidermolytic toxin by (1) biological tests in 3-day-old mice, (2) double-diffusion precipitation tests against specific antiserum, and (3) the appearance of characteristic protein bands on thin-layer-gel isoelectric focusing. Positive results were obtained in all three of these tests with supernates from 11 of these cultures; the same 11 strains, and no others, produced epidermal splitting when newborn mice were challenged with viable organisms. Of the 14 phage-group II strains included in the survey, eight (58%) produced epidermolytic toxin. Three toxinogenic strains belonged to phage groups other than group II. A radial-immunodiffusion test employing antiserum to purified epidermolytic toxin proved satisfactory for measuring amounts of epidermolytic toxin in excess of 200 mug per ml. The results of immunodiffusion tests indicated that six of the 11 positive strains produced two serologically distinct forms of epidermolytic toxin and that the remainder produced only one of these. A striking correlation was observed between the presence of toxin of serotypes i and ii and the occurrence of protein bands i and ii in thin-layer isoelectric-focusing gels. These tests should facilitate the qualitative and quantitative assessment of the production of different serotypes of epidermolytic toxin by S. aureus in future surveys of strains isolated from toxic epidermal necrolysis of Ritter's type and impetigo. The two forms of epidermolytic toxin previously designated ETA (pI=7-0) and ETB (pI=6-0) were detected by preparative isoelectric focusing of sero-type-i toxin. Evidence suggests that studies of the effect of heat should provide a means of investigating the relationship between the different molecular forms of epidermolytic toxin.