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Rapid identification of Candida albicans in blood by combined capillary electrophoresis and fluorescence in situ hybridization

机译：毛细管电泳-荧光原位杂交技术快速鉴定血液中的白色念珠菌

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摘要

A CE method based on whole-cell molecular labeling via fluorescence in situ hybridization was developed for the detection of Candida albicans in whole blood. Removal of potentially interfering red blood cells (RBC) with a simple hypotonic/detergent lysis step enabled us to detect and quantitate contaminating C. albicans cells at concentrations that were orders of magnitude lower than background RBC counts (∼7.0×109 RBC/mL). In the presence of the lysed blood matrix, yeast cells aggregated without the use of a blocking plug to stack the cells. Short (15 min) hybridizations yielded bright Candida-specific fluorescence in situ hybridization signals, enabling us to detect as few as a single injected cell. The peak area response of the stacked Candida cells showed a strong linear correlation with cell concentrations determined by plate counts, up to ∼107 CFU/mL (or ∼1×104 injected cells). This rapid and quantitative method for detecting Candida in blood may have advantageous applications in both human and veterinary diagnostics.
机译：建立了一种基于全细胞分子标记荧光原位杂交的CE方法，用于检测全血中的白色念珠菌。通过简单的低渗/去污裂解步骤去除潜在的干扰性红细胞（RBC），使我们能够检测和定量污染的白色念珠菌细胞，其浓度比背景RBC计数低几个数量级（〜7.0×10 9 RBC / mL）。在溶解的血液基质存在下，酵母细胞聚集而无需使用封闭塞来堆叠细胞。短时间（15分钟）的杂交产生明亮的念珠菌特异性荧光原位杂交信号，使我们能够检测到只有一个注射细胞的细胞。堆积的念珠菌细胞的峰面积响应与通过平板计数确定的细胞浓度呈强线性相关，高达〜10 7 CFU / mL（或〜1×10 4 注入的细胞）。这种用于检测血液中念珠菌的快速而定量的方法可能在人类和兽医诊断中都具有有利的应用。

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  • 来源
    《ELECTROPHORESIS》 |2010年第16期|p.2849-2853|共5页
  • 作者

    Andrew W. Lantz; Bledar Bisha; Man-Yung Tong; Ryan E. Nelson; Byron F. Brehm-Stecher; Daniel W. Armstrong;

  • 作者单位

    Department of Chemistry, Grand Valley State University, Allendale, MI, USA;

    Rapid Microbial Detection and Control Laboratory, Department of Food Science and Human Nutrition, Iowa State University, Ames, IA, USA;

    Department of Chemistry and Biochemistry, University of Texas, Arlington, TX, USA;

    Department of Chemistry, Grand Valley State University, Allendale, MI, USA;

    Rapid Microbial Detection and Control Laboratory, Department of Food Science and Human Nutrition, Iowa State University, Ames, IA, USA;

    Department of Chemistry and Biochemistry, University of Texas, Arlington, TX, USA;

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  • 原文格式 PDF
  • 正文语种 eng
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  • 关键词

    Blood; Candida albicans; CE; Fluorescence in situ hybridization;

    机译：血液;白色念珠菌;CE;荧光原位杂交;

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