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通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和免疫印迹分析对凝固酶阴性葡萄球菌进行鉴定

Characterization of coagulase-negative staphylococci by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analyses.

作者信息

Thomson-Carter F M, Pennington T H

机构信息

Department of Medical Microbiology, Medical School, Foresterhill, Aberdeen, Scotland, United Kingdom.

出版信息

J Clin Microbiol. 1989 Oct;27(10):2199-203. doi: 10.1128/jcm.27.10.2199-2203.1989.

Abstract

Coagulase-negative staphylococci are important nosocomial pathogens. At present, no wholly satisfactory typing scheme exists for these organisms. Therefore, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting were assessed as characterization methods. A total of 100 type strains and nontyped isolates representing nine species of coagulase-negative staphylococci were analyzed. Each species had a reproducible, characteristic whole-cell banding pattern when analyzed by either method. These species-specific profiles were obtained for all isolates despite disparate geographical origins and clinical isolation sites. Intraspecies similarities, calculated by using the Dice coefficient, were significantly higher than interspecies similarities. Although some species were more heterogeneous than others, the allocation of isolates to any particular species was reinforced by the high degree of interspecies dissimilarity. Application of SDS-PAGE also distinguished discrete subspecies groups. These groups possessed the characteristic profile of their species but were distinguished by a group of variable polypeptides. Species-specific banding patterns were also obtained by immunoblotting of whole-cell polypeptides. Differences between immunoblot and SDS-PAGE profiles could be attributed to variations of antigenicity of particular polypeptides. However, both SDS-PAGE and immunoblotting provided reproducible and sensitive methods for characterization of coagulase-negative staphylococci. Standardization of these techniques could provide the basis for a primary typing scheme.

摘要

凝固酶阴性葡萄球菌是重要的医院病原体。目前,尚无针对这些微生物的完全令人满意的分型方案。因此,评估了十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和免疫印迹作为鉴定方法。共分析了代表9种凝固酶阴性葡萄球菌的100株标准菌株和未分型分离株。通过这两种方法分析时,每个菌种都有可重复的、特征性的全细胞条带模式。尽管地理来源和临床分离部位不同,但所有分离株都获得了这些菌种特异性图谱。使用迪西系数计算的种内相似性显著高于种间相似性。虽然有些菌种比其他菌种更具异质性,但高度的种间差异加强了将分离株分配到任何特定菌种的准确性。SDS-PAGE的应用还区分了不同的亚种组。这些组具有其菌种的特征性图谱,但通过一组可变多肽得以区分。通过对全细胞多肽进行免疫印迹也获得了菌种特异性条带模式。免疫印迹和SDS-PAGE图谱之间的差异可归因于特定多肽抗原性的变化。然而,SDS-PAGE和免疫印迹都为凝固酶阴性葡萄球菌的鉴定提供了可重复且灵敏的方法。这些技术的标准化可为初步分型方案提供基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1167/266993/fa22655069fd/jcm00070-0068-a.jpg

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