UPMC Université Paris 06, UMR 7621, LOMIC, Observatoire Océanologique, Banyuls/mer, France.
J Appl Microbiol. 2010 Oct;109(4):1253-64. doi: 10.1111/j.1365-2672.2010.04752.x. Epub 2010 Aug 19.
We developed an improved Fluorescent In Situ Hybridization FISH-based method to detect viable Escherichia coli cells by solid phase cytometry (SPC), and results were compared to those obtained by the standard culture method.
The method includes a direct viable count (DVC) assay, multi-probes labelled and unlabelled (helpers) to detect specifically viable E. coli cells and to enhance SPC cell counts. We demonstrate that helpers increase the fluorescence intensity of hybridized E. coli cells as detected by SPC and assess the high specificity of the DVC-FISH procedure on a large panel of cultured strains. Application to seawater, freshwater and wastewater samples showed a good correlation between SPC cells counts and standard plate counts.
The high specificity of the procedure was demonstrated as well as its accuracy for detecting and counting viable E. coli cells in environmental samples.
The developed approach may be used to monitor faecal contamination sources and to investigate the occurrence of viable E. coli in natural environments.
我们开发了一种改良的荧光原位杂交（FISH）基于固相细胞术（SPC）的方法，用于检测活大肠杆菌细胞，并将结果与标准培养方法进行比较。
该方法包括直接活菌计数（DVC）测定，标记和未标记的多探针（辅助探针），以特异性检测活大肠杆菌细胞并增强 SPC 细胞计数。我们证明辅助探针可增强 SPC 检测到的杂交大肠杆菌细胞的荧光强度，并评估 DVC-FISH 程序在大量培养菌株上的高特异性。将其应用于海水、淡水和废水样本表明，SPC 细胞计数与平板计数之间具有良好的相关性。
该方法具有高度特异性，可准确检测和计数环境样本中的活大肠杆菌细胞。
该方法可用于监测粪便污染来源，并研究自然环境中活大肠杆菌的发生情况。
