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Rapid identification of Candida albicans in blood by combined capillary electrophoresis and fluorescence in situ hybridization
Authors:Andrew W Lantz  Bledar Bisha  Man‐Yung Tong  Ryan E Nelson  Byron F Brehm‐Stecher  Daniel W Armstrong
Abstract: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.
Keywords:Blood  Candida albicans  CE  Fluorescence in situ hybridization
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