Arbuthnott J P, Billcliffe B
J Med Microbiol. 1976 May;9(2):191-201. doi: 10.1099/00222615-9-2-191.
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.
通过以下方法对98株金黄色葡萄球菌培养物的浓缩上清液进行表皮溶解毒素产生情况的筛选：（1）在3日龄小鼠身上进行生物学试验；（2）针对特异性抗血清进行双向扩散沉淀试验；（3）在薄层凝胶等电聚焦上观察特征性蛋白条带的出现。在这98种培养物中，有11种培养物的上清液在上述所有三种试验中均得到阳性结果；同样是这11株菌株，而非其他菌株，在用活生物体攻击新生小鼠时会导致表皮分离。在所调查的14株噬菌体II组菌株中，有8株（58%）产生表皮溶解毒素。3株产毒素菌株属于除II组以外的其他噬菌体组。采用针对纯化表皮溶解毒素的抗血清的放射免疫扩散试验被证明可满意地测量每毫升超过200微克的表皮溶解毒素量。免疫扩散试验结果表明，11株阳性菌株中有6株产生两种血清学上不同形式的表皮溶解毒素，其余菌株仅产生其中一种。在薄层等电聚焦凝胶中，观察到血清型i和ii毒素的存在与蛋白条带i和ii的出现之间存在显著相关性。这些试验应有助于在未来对从里特氏型中毒性表皮坏死松解症和脓疱病分离的菌株进行的调查中，对金黄色葡萄球菌产生的不同血清型表皮溶解毒素进行定性和定量评估。通过血清型i毒素的制备性等电聚焦检测到了先前命名为ETA（pI = 7.0）和ETB（pI = 6.0）的两种表皮溶解毒素形式。有证据表明，对热效应的研究应提供一种手段来研究表皮溶解毒素不同分子形式之间的关系。
