一种用于快速检测和计数牛奶中总细菌数的流式细胞术方法。
A flow cytometry method for rapid detection and enumeration of total bacteria in milk.
作者信息
Gunasekera T S, Attfield P V, Veal D A
机构信息
Centre for Fluorimetric Applications in Biotechnology, Department of Biological Sciences, Macquarie University, Sydney, New South Wales 2109, Australia.
出版信息
Appl Environ Microbiol. 2000 Mar;66(3):1228-32. doi: 10.1128/AEM.66.3.1228-1232.2000.
Abstract
Application of flow cytometry (FCM) to microbial analysis of milk is hampered by the presence of milk proteins and lipid particles. Here we report on the development of a rapid (</=1-h) FCM assay based on enzymatic clearing of milk to determine total bacteria in milk. When bacteria were added to ultra-heat-treated milk, a good correlation (r >/= 0.98) between the FCM assay and the more conventional methods of plating and direct microscopic counting was achieved. Raw milk data showed a significant correlation (P < 0.01) and a good agreement (r = 0.91) between FCM and standard plate count methods. The detection limit of the FCM assay was </=10(4) bacteria ml of milk(-1). This limit is below the level of detection required to satisfy legislation in many countries and states.
摘要
牛奶中的蛋白质和脂质颗粒会妨碍流式细胞术（FCM）在牛奶微生物分析中的应用。在此，我们报告一种基于酶解牛奶的快速（≤1小时）FCM检测方法的开发，用于测定牛奶中的总细菌数。当将细菌添加到超高温处理的牛奶中时，FCM检测方法与更传统的平板计数法和直接显微镜计数法之间具有良好的相关性（r≥0.98）。生鲜牛奶数据显示，FCM与标准平板计数法之间具有显著相关性（P<0.01）和良好的一致性（r = 0.91）。FCM检测方法的检测限≤10⁴ 个细菌/毫升牛奶。这个限值低于许多国家和州的立法要求的检测水平。
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