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快速检测单细胞细菌作为食品微生物学中的一种新方法。

Rapid detection of single cell bacteria as a novel approach in food microbiology.

作者信息

D'Haese Eva, Nelis Hans J

机构信息

Ghent University, Laboratory for Pharmaceutical Microbiology, Belgium.

出版信息

J AOAC Int. 2002 Jul-Aug;85(4):979-83.

Abstract

Solid-phase cytometry (SPC) is a novel technique that allows rapid detection of bacteria at the single cell level, without the need for a growth phase. After filtration of the sample, the retained microorganisms are fluorescently labeled on the membrane filter and automatically counted by a laser scanning device. Each fluorescent spot can be visually inspected with an epifluorescence microscope connected to the ChemScan by a computer-driven moving stage. Depending on the fluorogenic labels used, information on the identity and the physiological status of the microorganisms can be obtained within a few hours. Although SPC was originally recommended for the determination of the total viable microbial count in water and other liquid samples, it may also be a promising technique for the detection and enumeration of bacteria in food samples, provided they can be isolated from the unfilterable matrix. The short detection time inherent in this approach is a considerable advantage over conventional plate counting, especially for slow-growing microorganisms. The basic principles of SPC are discussed as well as its potential for the detection of Mycobacterium paratuberculosis, a model example of a slow-growing bacterium in milk.

摘要

固相细胞术(SPC)是一项新技术,可在单细胞水平快速检测细菌,无需细菌处于生长阶段。样品过滤后,截留的微生物在膜滤器上进行荧光标记,并通过激光扫描装置自动计数。每个荧光点可通过连接到ChemScan的落射荧光显微镜,利用计算机驱动的移动载物台进行目视检查。根据所使用的荧光标记,可在数小时内获得有关微生物的身份和生理状态的信息。尽管SPC最初推荐用于测定水和其他液体样品中的总活微生物数,但对于食品样品中的细菌检测和计数,它也可能是一项有前景的技术,前提是它们可以从未过滤的基质中分离出来。这种方法固有的短检测时间是相对于传统平板计数法的一个显著优势,特别是对于生长缓慢的微生物。本文讨论了SPC的基本原理及其检测副结核分枝杆菌的潜力,副结核分枝杆菌是牛奶中生长缓慢细菌的一个典型例子。

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