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假丝酵母“糙球拟内孢霉”——电迁移技术和 MALDI-TOF 质谱用于表型鉴别。

Candida "psilosis"--electromigration techniques and MALDI-TOF mass spectrometry for phenotypical discrimination.

机构信息

Institute of Analytical Chemistry of the ASCR, v. v. i., Veveří 97, 602 00 Brno, Czech Republic.

出版信息

Analyst. 2012 Apr 21;137(8):1937-43. doi: 10.1039/c2an15931g. Epub 2012 Mar 7.

Abstract

Biofilm-positive strains of Candida parapsilosis are the second most common yeasts responsible for bloodstream infections. This pathogen is difficult to identify by standard methods from other phenotypically indistinguishable species, biofilm-negative Candida orthopsilosis and biofilm-positive Candida metapsilosis. From a medical point of view, important information is especially whether the strains form biofilm. The biofilm formation enables yeast to colonize artificial surfaces thereby protecting the yeast cell against antifungal agents. The commonly used genotypic methods including different modifications of the polymerase chain reaction have some disadvantages. Therefore, a rapid and reliable method able to identify phenotypically indistinguishable C. "psilosis" species is still of great interest. In this study, the four well-established analytical techniques: gel isoelectric focusing, sodium dodecyl sulfate polyacrylamide gel electrophoresis, two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, were applied in order to discriminate C. "psilosis" species. The ability of these techniques to differentiate between biofilm-positive and biofilm-negative strains was further investigated. Our results have revealed that the proposed methods, especially matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of intact yeast cells, can be used as the efficient tools for discrimination and identification of the phenotypically indistinguishable microorganisms.

摘要

假丝酵母中生物膜阳性菌株是导致血流感染的第二大常见酵母。与其他表型难以区分的物种(生物膜阴性近平滑假丝酵母和生物膜阳性中间假丝酵母)相比,这种病原体很难通过标准方法识别。从医学角度来看,重要的信息是菌株是否形成生物膜。生物膜的形成使酵母能够在人工表面定植,从而保护酵母细胞免受抗真菌药物的侵害。常用的基因分型方法包括聚合酶链反应的不同修饰,都存在一些缺点。因此,能够识别表型难以区分的 C. “假丝酵母”物种的快速可靠方法仍然具有重要意义。在这项研究中,应用了四种成熟的分析技术:凝胶等电聚焦、十二烷基硫酸钠聚丙烯酰胺凝胶电泳、二维凝胶电泳和基质辅助激光解吸/电离飞行时间质谱,以区分 C. “假丝酵母”物种。进一步研究了这些技术区分生物膜阳性和生物膜阴性菌株的能力。我们的研究结果表明,所提出的方法,尤其是完整酵母细胞的基质辅助激光解吸/电离飞行时间质谱,可以作为区分和鉴定表型难以区分的微生物的有效工具。

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