Bayston R, Rodgers J
Department of Paediatric Surgery, Institute of Child Health, London.
J Clin Pathol. 1990 Oct;43(10):866-70. doi: 10.1136/jcp.43.10.866.
A method of optimising slime production produced by Staphylococcus epidermis and its quantitative assay was developed, which gave a preliminary indication of its identity and an assessment of the correlation between slime production and adherence of the organism to implants. After inducing vigorous growth in brain heart infusion broth to stationary phase, all nutrients were removed by washing and the organisms resuspended in sterile deionised water with added magnesium. After further incubation the culture was centrifuged and the supernatant reacted with alcian blue in 50 mM magnesium chloride/sodium acetate solution, and the amount of bound dye was measured spectrophotometrically at 620 nm after its resolubilisation using sodium dodecyl sulphate. Large quantities of slime were produced by some, but not all, strains. Preliminary electrophoresis of the slime showed mobility and staining similar to that of the glycosaminoglycans. Adherence was tested by growing strains in wells of tissue culture plates and aspirating the supernatant after incubation. After fixation and staining of adherent growth the amount of bound stain was determined spectrophotometrically after its elution with ethanol. In this series of organisms there was no correlation between the result of tests for adherence or production of extracellular slime, and no correlation between either of these and the clinical source of the organisms.
开发了一种优化表皮葡萄球菌产生的黏液产量及其定量测定的方法,该方法初步表明了其特性,并评估了黏液产量与该生物体在植入物上黏附之间的相关性。在脑心浸液肉汤中诱导旺盛生长至稳定期后,通过洗涤去除所有营养物质,将生物体重悬于添加了镁的无菌去离子水中。进一步孵育后,将培养物离心,上清液与在50 mM氯化镁/醋酸钠溶液中的阿尔新蓝反应,使用十二烷基硫酸钠使其重新溶解后,在620 nm处用分光光度法测量结合染料的量。一些但并非所有菌株都产生了大量黏液。黏液的初步电泳显示其迁移率和染色与糖胺聚糖相似。通过在组织培养板的孔中培养菌株并在孵育后吸出上清液来测试黏附性。在固定并染色黏附生长物后,用乙醇洗脱后用分光光度法测定结合染料的量。在这一系列生物体中,黏附测试结果或细胞外黏液产生结果之间没有相关性,并且这些结果与生物体临床来源之间也没有相关性。