[1]张帅,齐颖颖,张红星,等.快速检测生鲜肉中三种食源性致病菌[J].江苏农业学报,2016,(04):939-945.[doi:10.3969/j.issn.100-4440.2016.04.036]
　ZHANG Shuai,QI Ying-ying,ZHANG Hong-xing,et al.Rapid detection of three foodborne pathogens in raw meat[J].,2016,(04):939-945.[doi:10.3969/j.issn.100-4440.2016.04.036]
点击复制

快速检测生鲜肉中三种食源性致病菌()
分享到：

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2016年04期
页码:
939-945
栏目:
加工贮藏·质量安全
出版日期:
2016-10-31

文章信息/Info

Title:
Rapid detection of three foodborne pathogens in raw meat
作者:
张帅; 齐颖颖; 张红星; 王怡雯; 谢远红; 刘慧
北京农学院食品科学与工程学院/食品质量与安全北京实验室/农产品有害微生物及农残安全检测与控制北京市重点实验室/北京市食品安全免疫快速检测工程技术研究中心/微生态制剂关键技术开发北京市工程实验室，北京 102206
Author(s):
ZHANG Shuai; QI Ying-ying; ZHANG Hong-xing; WANG Yi-wen; XIE Yuan-hong; LIU-Hui
Beijing Laboratory of Food Quality and Safety/Beijing Key Laboratory of Agricultural Product Detection and Control for Spoilage Organisms and Pesticide/Beijing Engineering Technology Research Center of Food Safety Immune Rapid Detection/Beijing Engineering Laboratory of Key Technology Development of Microecologics/College of Food Science and Engineering, Beijing University of Agriculture, Beijing 102206，China
关键词:
食源性致病菌; 单克隆抗体; 多克隆抗体; 胶体金免疫层析试纸条
Keywords:
foodborne pathogen;  monoclonal antibody;  polyclonal antibody;  immunochromatographic strip
分类号:
S851.347.02
DOI:
10.3969/j.issn.100-4440.2016.04.036
文献标志码:
A
摘要:
为同时快速检测生鲜肉中鼠伤寒沙门氏菌、单增李斯特菌和福氏志贺氏菌3种食源性致病菌，降低检测成本，制备了特异性单克隆抗体和多克隆抗体，研制出能够同时检测以上3种致病菌的胶体金免疫层析试纸条。结果表明，本试验成功筛选出3株稳定分泌抗体的杂交瘤细胞株2E3、2E7和2E8，3种单克隆抗体效价均在1.28×106以上，3种多克隆抗体效价均在2.00×106以上。制备的胶体金免疫层析试纸条对鼠伤寒沙门氏菌、单增李斯特菌和福氏志贺氏菌3种食源性致病菌的特异性良好，灵敏度均为1×104CFU/g。
Abstract:
To achieve the simutaneous detection of Salmonella typhimurium, Listeria monocytogenes and Shigella flexneri in raw meat, three hybridoma cell strains, 2E3, 2E7 and 2E8, stably secreting antibodies against above pathogens were screened. The titers of three monoclonal antibodies and three polyclonal antibodies were more than 1.28×106 and 2.00×106, respectively. The immunochromatographic strip developed could detect all three pathogens as low as 1×104 CFU/g. L. monocytogenes was detected by the strip in market sample which was free of other two pathogens.

参考文献/References:

[1]蒲承君, 崔兴博, 杜兰, 等. 市售生鲜鸡肉5种主要食源性致病菌污染检测及分布概率评估[J]. 食品科学,2015,36(10): 201-205.
[2]CALLAWAY T R, EDRINGTON T S, ANDERSON R C, et al. Gastrointestinal microbial ecology and the safety of our food supply as related to Salmonella[J]. Journal of Animal Science, 2008, 86 (14):163-172. 
[3]JIN H, CHETAN J, LEE J H. Antimicrobial resistance of Salmonella isolated from food animals: a review[J]. Food Research International,2012, 45(2): 819-830.
[4]宫强, 李战丽, 牛明福, 等. 鼠伤寒沙门氏菌ompc基因PCR的检测方法[J].食品科学,2015,36 (16): 251-254.
[5]颜成英，汤晓艳，康大成，等.EMA PTPCR法检测鸡肉中鼠伤寒沙门氏菌活菌[J].中国食品学报,2015,15(10):179-184.
[6]SHOUKAT S, MALIK S V S, RAWOOL D B, et al. A study on detection of pathogenic Listeria monocytogenes in ovine’s of kashmir region faving abortion or history of abortion[J]. Proceedings of the National Academy of Sciences,India Section B:Biological Sciences,2014,84(2):311-316.
[7]李翠云. 单核细胞李斯特菌研究近况[J]. 中国热带医学,2010(1): 12-122.
[8]VAZQUEZ-BOLAND J A, KUHN M, BERCHE P, et al. Listeria pathogenesis and molecular virulence determinants [J].Clinical Microbiology Reviews, 2001, 14(3): 584-640.
[9]CABANES D, DEHOUX P, DUSSURGET O, et al. Surface proteins and the pathogenic potential of Listeria monocytogenes [J].Trends in Microbiology, 2002, 10(5): 238-245.
[10] WARREN B R, PARISH M E, SCHNEIDER K R. Shigella as a food borne pathogen and current methods for detection in food[J] .Critical Review s in Food Science and Nutrition, 2006, 46(7):551-567.
[11] 陈伟, 李正国, 杨平, 等.肉制品中志贺氏菌DNA的快速提取方法[J]. 食品发酵工业,2009,35(5):257-261.
[12] 任娟.胶体金免疫层析技术研究现状及进展[J].新疆畜牧业,2011(12): 22-24.
[13] 徐超莲, 赖卫华, 刘道峰.胶体金免疫层析法检测食品中天然存在的危害物质的研究进展[J].食品科学, 2014, 35(5): 257-261.
[14] 郭建军.胶体金免疫层析技术在水产品药物残留检测中的应用[J].北京农业, 2014(5): 7-10.
[15] MOONGKARNDI P, RODPAI E, KANARAT S. Evaluation of an immunochromatographic assay for rapid detection of Salmonella enterica serovars Typhimurium and enteritidis[J]. Journal of Veterinary Diagostic Investigation, 2011,23(4): 797-801. 
[16] 伍燕华, 牛瑞江, 赖卫华, 等. 双抗夹心酶联免疫吸附法检测沙门氏菌[J]. 食品工业科技, 2014, 35(10): 62-65.
[17] 夏俊芳, 刘芳, 刘箐, 等.可视化抗体阵列快速联合检测大肠杆菌O157∶H7 和鼠伤寒沙门氏菌[J]. 微生物学通报, 2014,41(7): 1382-1393.
[18] 兰全学, 刘小青, 李幸桓, 等. 多重PCR联合同源加尾技术检测四种食源性致病菌[J]. 中国食品工业,2014(6): 59-64.
[19] LIN M, TODORIC D, MALLORY M, et al. Monoclonal antibodies binding to the cell surface of Listeria monocytogenes serotype 4b[J].Journal of Medical Microbiology,2006,55: 291-299．
[20] 霍华德 G C, 凯瑟 M R.抗体制备与使用实验指南[M].北京:科学出版社,2010:97.
[21] 牛瑞江, 赖卫华, 熊齐荣, 等.鼠伤寒沙门氏菌特异性多克隆抗体的纯化方法[J]. 食品科学,2011,32(13):151-155.
[22] 李任峰.检测口蹄疫病毒单抗ELISA方法的建立及口蹄疫病毒多抗原基因的融合表达[D].武汉：华中农业大学,2005.
[23] 齐颖颖, 吴萌, 王怡雯, 等,单增李斯特菌单克隆抗体的研制及特性分析[J].食品与生物技术学报,2016,35(2):151-155.
[24] 齐颖颖.单增李斯特菌的单克隆抗体制备及胶体金试纸条的研制[D].北京：北京农学院,2014.
[25] 卫生部,国家标准化管理委员会.食品微生物学检验:沙门氏菌检验：GB 4789.4-2010[S].北京:中国标准出版社,2010:25-44．
[26] 卫生部,国家标准化管理委员会.食品微生物学检验:单核细胞增生李斯特氏菌检验：GB 4789.3-2010[S]．北京:中国标准出版社,2010:299-308．
[27] 卫生部,国家标准化管理委员会.食品微生物学检验:志贺氏菌检验：GB 4789.5-2012[S]. 北京:中国标准出版社,2012:45-60．
[28] 夏诗琪,徐超莲,刘道峰,等.胶体金免疫层析法联检食品中5种典型沙门氏菌模型的建立和优化[J].食品科学,2014,35（22）:154-158.

相似文献/References:

[1]王怡雯,齐颖颖,张红星,等.福氏志贺氏菌2a单克隆抗体的制备及ELISA方法的建立[J].江苏农业学报,2017,(02):450.[doi:doi:10.3969/j.issn.1000-4440.2017.02.033]
　WANG Yi-wen,QI Ying-ying,ZHANG Hong-xing,et al.Preparation of Shigella flexneri 2a monoclonal antibodies and establishment of ELISA for its detection[J].,2017,(04):450.[doi:doi:10.3969/j.issn.1000-4440.2017.02.033]
[2]李盼,张晓帅,邱雨楼,等.有机磷农药广谱特异性单克隆抗体的制备及特性分析[J].江苏农业学报,2017,(06):1415.[doi:doi:10.3969/j.issn.1000-4440.2017.06.031]
　LI Pan,ZHANG Xiao-shuai,QIU Yu-lou,et al.Production and characterization of broad-specific monoclonal antibody for organophosphorus pesticides recognization[J].,2017,(04):1415.[doi:doi:10.3969/j.issn.1000-4440.2017.06.031]
[3]何方洋,杨昌松,韩深,等.应用酶联免疫法测定环境水样中雌三醇的残留量[J].江苏农业学报,2017,(06):1422.[doi:doi:10.3969/j.issn.1000-4440.2017.06.032]
　HE Fang-yang,YANG Chang-song,HAN Shen,et al.Enzyme-linked immunosorbent assay（ELISA） for determination of estriol in environmental water sample[J].,2017,(04):1422.[doi:doi:10.3969/j.issn.1000-4440.2017.06.032]
[4]任春梅,杨柳,缪倩,等.小麦黄花叶病毒单克隆抗体的制备及ACP-ELISA检测方法的建立[J].江苏农业学报,2018,(01):34.[doi:doi:10.3969/j.issn.1000-4440.2018.01.005]
　REN Chun-mei,YANG Liu,MIAO Qian,et al.Production of monoclonal antibodies to wheat yellow mosaic virus and ACP-ELISA application in virus detection[J].,2018,(04):34.[doi:doi:10.3969/j.issn.1000-4440.2018.01.005]
[5]祁光宇,刘斌,赵兴绪.猪流行性腹泻病毒HB2015分离株N基因表达及单克隆抗体的制备[J].江苏农业学报,2018,(01):76.[doi:doi:10.3969/j.issn.1000-4440.2018.01.011]
　QI Guang-yu,LIU Bin,ZHAO Xing-xu.Expression of N gene of porcine epidemic diarrhea virus（PEDV） HB2015 strain and preparation of monoclonal antibodies against PEDV[J].,2018,(04):76.[doi:doi:10.3969/j.issn.1000-4440.2018.01.011]
[6]杨柳,任春梅,缪倩,等.黄瓜绿斑驳花叶病毒的双抗夹心ELISA检测[J].江苏农业学报,2018,(04):769.[doi:doi:10.3969/j.issn.1000-4440.2018.04.008]
　YANG Liu,REN Chun-mei,MIAO Qian,et al.DAS-ELISA detection against cucumber green mottle mosaic virus[J].,2018,(04):769.[doi:doi:10.3969/j.issn.1000-4440.2018.04.008]

备注/Memo

备注/Memo:
收稿日期：2016-05-20基金项目：北京市属高等学校高层次人才引进与培养长城学者计划（CIT&TCD20140315）；北京农学院协同创新建设项目作者简介：张帅(1992-)，男，山东聊城人，硕士研究生，主要从事食品安全快速检测研究。 (E-mail)15701299950@163.com通讯作者：张红星，(E-mail)hxzhang511@163.com
更新日期/Last Update: 2016-11-01