广西师范大学学报（自然科学版） ›› 2017, Vol. 35 ›› Issue (2): 112-116.doi: 10.16088/j.issn.1001-6600.2017.02.017

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流式细胞术对生乳中微生物检测的应用研究

杨莉婷1, 2, 何丽2, 何海宁1, 吴琼1, 2*   

  1. 1.甘肃省动物源性制品安全分析与检测技术重点实验室,甘肃兰州730000;
    2.广西师范大学生命科学学院,广西桂林541006
  • 出版日期:2017-07-25 发布日期:2018-07-25
  • 通讯作者: 吴琼(1984—),女,甘肃静宁人,广西师范大学副教授,博士。E-mail:wubinbin02@163.com
  • 基金资助:
    国家自然科学基金(31660344);甘肃省动物源性制品安全分析与检测技术重点实验室开放课题(2015-2);广西大学生创新创业训练项目(201610602052)

The Application of Flow Cytometry in Testing Bacteria of Raw Milk

YANG Liting1,2, HE Li2, HE Haining1, WU Qiong1,2*   

  1. 1. Key Laboratory of Animal Products Safety Analysis and Detection Technology of Gansu Province, Lanzhou Ganshu730000, China;
    2. College of Life Science, Guangxi Normal University, Guilin Guangxi 541006, China
  • Online:2017-07-25 Published:2018-07-25

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摘要： 流式细胞术(flow cytometry,FCM)是一种对流动的单细胞或小颗粒分子进行快速、多参数检测的新兴技术,其以快速、精确的特点,被应用于食品微生物的检测中。本研究以牛乳为材料,使用蛋白酶消化和尼龙膜过滤的方法来去除生乳中的大颗粒物质;利用7-AAD进行染色,结合荧光微球,对细菌总数进行计数;在FACS Aria II flow cytometer (BD)流式细胞仪的操作系统上建立微生物的检测程序检测细菌总数;检测限达到101 CFU/mL 数量级。检测结果与平皿菌落计数法进行比较分析,结果表明,流式细胞术与平皿菌落计数法的结果呈显著相关(P<0.01)。

关键词: 流式细胞术, 食品检测, 微生物, 牛乳

Abstract: Flow cytometry (FCM) is a rapid technique for cell-by-cell multiparameter analysis, which is often used in combination with fluorescent labelling for fast viability assessment. FCM analysss with fast and accurate protocol appeared to be workable for food bacteria detection. This article applied enzymatic treatment and membrane filtration method to digest proteins and treatment of milk by centrifugation to remove lipids. Treated milk samples were detected and enumerated by FCM. The bacteria were stained with 7-AAD and counted by fluorescent microspheres. The detection limit of 101 CFU/mL was achieved. Raw milk data showed a significant correlation (P<0.01) and a good agreement(r=0.980 2) between FCM and standard plate count methods.

Key words: flow cytometry, food detection, bacteria, milk

中图分类号: 

  • Q932
[1] 柴阿丽,韩云,武军,等.基于FDA-PI双荧光复染法的茄病镰刀菌孢子活性检测[J].中国农业科学, 2015,48(14):2757-2766.
[2] GAN Nanqin, HUANG Qun,ZHENG Lingling,et al.Quantitative assessment of toxic and nontoxic Microcystis colonies in natural environments using fluorescence in situ hybridization and flow cytometry[J].Science China(Life Sciences),2010,53(8):973-980.
[3] ELHADIDY A M,VAN DYKE M I,PELDSZUS S,et al. Application of flow cytometry to monitor assimilable organic carbon (AOC) and microbial community changes in water[J]. Journal Microbiol Methods, 2016,9(13):9-18.
[4] LANGERHUUS S N,INGVARTSEN K L,BENNEDSGAARD T W,et al. Gram-typing of mastitis bacteria in milk samples using flow cytometry[J]. Journal of Dairy Science,2013,96(1):267-277.
[5] HILDEBRANDT P,SURMANN K,SALAZAR M G,et al. Alternative fluorescent labeling strategies for characterizing gram-positive pathogenic bacteria: flow cytometry supported counting, sorting, and proteome analysis of Staphylococcus aureus retrieved from infected host cells[J]. Cytometry Part A, 2016,9(19):1-9.
[6] 王宁,刘宁.流式细胞术快速检测生乳中细菌总数[J].食品工业科技,2007,28(9):197-200.
[7] 刘道亮,赵占民,胡连霞,等.应用流式细胞技术快速检测液态商品中的细菌总数[J].食品科学,2011,32(2):157-163.
[8] 林怡雯,杨天,李丹,等.基于CTC-流式细胞仪活性细菌总数的快速检测技术研究[J].环境科学学报,2013,33(9):2511-2515.
[9] 李影.大肠杆菌、乳酸杆菌“活的非可培养状态”研究[D]. 长春:吉林农业大学,2007.
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