摘要
建立一种基于流式分析技术的快速定量检测牛乳中大肠杆菌O157:H7的方法。用偶联有异硫氰酸荧光素的大肠杆菌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(phosphate buffer saline,PBS)和人工污染牛乳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在PBS中的检测范围为2.57×10~3~1.12×10~8 CFU/m L,灵敏度达到2.57×10~3 CFU/m L。将所建立的流式检测方法应用于牛乳样品检测,当人工污染牛乳样品中大肠杆菌O157:H7的浓度在2.31×10~4~1.48×10~8 CFU/m L之间时,流式检测方法与平板计数方法检测结果基本一致,方法的灵敏度为2.31×10~4 CFU/m L,检测时间为35 min。该方法能快速、定量地检测出牛乳样品中的大肠杆菌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。
The purpose of this study was to develop a flow cytometry(FCM)method for rapid and quantitative detection of Escherichia coli O157:H7in milk.E.coli O157:H7was labeled with fluorescein isothiocyanate(FITC)-conjugated anti-E.coli O157monocolonal antibodies and then detected by flow cytometry(green fluorescence channel).Serial10-fold dilutions of E.coli O157:H7in PBS were tested by FCM method under optimized reaction conditions.The detection range of this method was2.57×103–1.12×108CFU/mL and the sensitivity was2.57×103CFU/mL.Then the FCM method was applied to artificially contaminated milk samples with2.31×104–1.48×108CFU/mL E.coli O157:H7.The results were consistent with the plate counting method.The sensitivity of our method was2.31×104CFU/mL and the process required35min.This method allowed for rapid,efficient and sensitive detection of E.coli O157:H7in milk samples.Thus,it has a great potential for application in rapid screening and monitoring of foodborne pathogens.
作者
刘思渊
古少鹏
隋志伟
刘思章
薛蕾
霍乃蕊
LIU Siyuan;GU Shaopeng;SUI Zhiwei;LIU Sizhang;XUE Lei;HUO Nairui(College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China;National Institute of Metrology, Beijing 100013, China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2018年第6期302-306,共5页
Food Science
基金
山西省重点研发计划重点项目(201603D21108-03)
"十三五"国家重点研发计划重点专项(2017YFF0204602)
国家标准物质资源共享平台项目(32-APT1702-9)
