Comparative assessment of epifluorescence microscopy, flow cytometry and solid-phase cytometry used in the enumeration of specific bacteria in water
Comparative assessment of epifluorescence microscopy, flow cytometry and solid-phase cytometry used in the enumeration of specific bacteria in water
复制标题
落射荧光显微镜、流式细胞术和固相细胞术在水中特定细菌计数中的比较评估
DOI：
10.3354/ame025301
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发表时间：
2001
影响因子：
1.4
通讯作者：
P. Lebaron
中科院分区：
文献类型：
--
作者：
K. Lemarchand;N. Parthuisot;P. Catala;P. Lebaron
快速微生物学和罕见事件的检测是水生微生物学各个领域的重要挑战。落射荧光显微术、流式细胞术和固相细胞测定法是用于微生物学中直接方法的技术，但这些不同仪器的应用范围未明确定义。在这项研究中，我们检查了每种技术可以可靠使用的细菌浓度下限。比较了（1）荧光珠、（2）不同标记/未标记细胞比例的标记细菌、（3）自来水中不同密度接种的大肠杆菌O 157：H7细胞和（4）大肠杆菌的Enumeration技术。大肠杆菌O 157：H7细胞。尽管存在显著的未标记细胞背景，但不同方法得到的结果相关性良好。然而，固相细胞术是允许精确计数罕见事件（低至1个细胞）的唯一技术，其提供与传统培养方法相同的灵敏度。检测灵敏度不受高达10 7未标记细胞的过滤器上的存在。相比之下，流式细胞术是一种非常快速和准确的方法，但它不能应用于罕见事件的检测。E.大肠杆菌O 157：H7细胞可在非特异性细菌存在的环境水样中快速、准确地检测出来。固相细胞仪结合分类探针可以快速准确地检测各种各样的水生环境中的生态利益的种类繁多。
Rapid microbiology and the detection of rare events are important challenges in various fields of aquatic microbiology. Epifluorescence microscopy, flow cytometry and solid-phase cyto- metry are techniques used for direct methods in microbiology but the range of application of these different instruments is not clearly defined. In this study, we examined the lower limit of bacterial concentration to which each technique can be reliably used. Techniques were compared for the enu- meration of (1) fluorescent beads, (2) labeled bacteria at different ratios of labeled/non-labeled cells, (3) Escherichia coli O157:H7 cells inoculated at different densities in tap water, and (4) E. coli O157:H7 cells in artificially contaminated natural seawater. The different methods gave results that correlated well despite the presence of a significant background of unlabeled cells. However, solid- phase cytometry was the only technique that allowed the accurate enumeration of rare events (down to 1 cell) providing the same sensitivity as traditional culture methods. The detection sensitivity was not affected by the presence of up to 10 7 unlabeled cells on the filter. In contrast, flow cytometry was a very rapid and accurate method but it could not be applied to the detection of rare events. E. coli O157:H7 cells could be detected rapidly and accurately in environmental water samples in the pres- ence of non-specific bacteria. Solid-phase cytometry combined with taxonomic probes allowed rapid and accurate detection of a large variety of species of ecological interest in a wide variety of aquatic environments.