基于流式分析技术的牛奶中大肠杆菌O157:H7快速定量检测

摘要/Abstract

摘要： 本研究的目的是建立一种基于流式分析技术的快速定量检测牛奶中大肠杆菌O157:H7的新方法。用偶联有异硫氰酸荧光素（FITC）的大肠杆菌O157单克隆抗体对大肠杆菌O157:H7进行特异性标记，通过优化抗体反应条件，建立流式检测方法，然后对磷酸盐缓冲溶液和人工污染牛奶样品中不同浓度的大肠杆菌O157:H7进行定量检测。本研究建立的流式检测方法的检测范围为2.57 × 103-1.12 × 108 CFU/mL，灵敏度达到2.57 × 103 CFU/mL。将所建立的流式检测方法应用于牛奶样品检测，当人工污染牛奶样品中大肠杆菌O157:H7的浓度在2.31 × 104-1.48 × 108 CFU/mL范围时，流式检测方法与平板计数方法检测结果基本一致，流式检测方法的灵敏度为2.31 × 104 CFU/mL，检测时间为35 min。该方法能快速、高效、灵敏地检测出牛奶样品中的大肠杆菌O157:H7，在食源性致病菌的快速筛查和监控中具有重要的应用价值。

关键词: 流式分析技术, 快速检测, 大肠杆菌O157:H7, 牛奶

Abstract: The purpose of this study is to develop a flow cytometry (FCM)-based method for rapid and quantitative detection of Escherichia coli O157:H7 in milk. In this study, Escherichia coli O157:H7 was labeled with FITC by reaction with the anti- Escherichia coli O157 monoclonal antibody(FITC) and then dectected by flow cytometry (green fluorescence channel). Serial 10-fold dilution of Escherichia coli O157:H7 in PBS was tested by FCM method with optimized reaction conditions, the detection range of this method is 2.57×103-1.12 ×108 CFU/mL and the sensitivity is 2.57 × 103 CFU/mL. Then the FCM method was applied to the detection of milk samples. When the concentration of Escherichia coli O157:H7 in artificially contaminated milk samples is between 2.31 × 104-1.48 × 108 CFU/mL, the results of FCM method is consistent with the plate counting method. The sensitivity of this method is 2.31 × 104 CFU/mL and total time is 35 min. This method can detect Escherichia coli O157:H7 in milk samples rapidly, efficiently and sensitively. So it has a great application value in rapid screening and monitoring of foodborne pathogens.

Key words: Flow cytometry technology, Rapid detection, Escherichia coli O157:H7, Milk

中图分类号:

TS207.4

刘思渊古少鹏隋志伟刘思章薛蕾霍乃蕊. 基于流式分析技术的牛奶中大肠杆菌O157:H7快速定量检测[J]. 食品科学, 0, (): 0-0.

Shaopeng Shao.P.GuGu Zhi-Wei SUI Nairui Nai.R.HuoHuo. Rapid and Quantitative Detection of Escherichia coli O157:H7 in Milk Based on Flow Cytometry Technology[J]. FOOD SCIENCE, 0, (): 0-0.

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