Abstract: Multiplex PCR technology is developed on the basis of conventional PCR technology,its reaction principle,reaction reagent and operation process are the same as conventional PCR.The difference is that multiplex PCR technology adds two or more pairs of primers in the same reaction system to amplify different templates and get different target fragments.Multiplex PCR technology has the advantages of high throughput and cost saving in the detection of foodborne pathogens,so it has a good development prospect in the actual detection.This paper summarizes the research progress of multiplex PCR technology in the detection of foodborne pathogens and its current situation in standard formulation and commercial application,and puts forward the technical points and future research directions of multiplex PCR technology in the detection of foodborne pathogens.
[1] 赵彤.食源性致病菌检测现状与食品微生物危险性评估[J].中国卫生标准管理,2019,10(4):7-9. ZHAO T.Detection status of food borne pathogenic bacteria and food microbial risk assessment[J].China Health Standard Management,2019,10(4):7-9. [2] 中华人民共和国国家卫生和计划生育委员会,国家食品药品监督管理总局.GB 4789.4—2016 食品安全国家标准 食品微生物学检验 沙门氏菌检验[S].北京:中国标准出版社,2016. National Health and Family Planning Commission of the People′s Republic of China,China Food and Drug Administration.GB 4789.4—2016 National food safety standard Food microbiological examination:Examination of Salmonella[S].Beijing:China Standards Press,2016. [3] 中华人民共和国国家卫生部.GB 4789.5—2012食品安全国家标准 食品微生物学检验 志贺氏菌检验[S].北京:中国标准出版社,2012. Ministry of Health of the People′s Republic of China.GB 4789.5—2012 National food safety standard Food microbiological examination:Examination of Shigella[S].Beijing:China Standards Press.2012. [4] 中华人民共和国国家卫生和计划生育委员会,国家食品药品监督管理总局.GB 4789.6—2016 食品安全国家标准 食品微生物学检验 致泻大肠埃希氏菌检验[S].北京:中国标准出版社,2016. National Health and Family Planning Commission of the People′s Republic of China,China Food and Drug Administration.GB 4789.6—2016 National food safety standard Food microbiological examination:Examination of diarrheagenic Escherichia coli[S].Beijing:China Standards Press.2016. [5] 中华人民共和国国家卫生和计划生育委员会.GB 4789.7—2013 食品安全国家标准 食品微生物学检验 副溶血性弧菌检验[S].北京:中国标准出版社,2013. National Health and Family Planning Commission of the People′s Republic of China.GB 4789.7—2013 National food safety standard Food microbiological examination:Examination of Vibrio parahaemolyticus[S].Beijing:China Standards Press.2013. [6] 中华人民共和国国家卫生和计划生育委员会.GB 4789.8—2016 食品安全国家标准 食品微生物学检验 小肠结肠炎耶尔森氏菌检验[S].北京:中国标准出版社,2016. National Health and Family Planning Commission of the People′s Republic of China.GB 4789.8—2016 National food safety standard Food microbiological examination:Examination of Yersinia enterocolitica[S].Beijing:China Standards Press,2016. [7] 中华人民共和国国家卫生和计划生育委员会.GB 4789.9—2014 食品安全国家标准 食品微生物学检验 空肠弯曲菌检验[S].北京:中国标准出版社.2014. National Health and Family Planning Commission of the People′s Republic of China.GB 4789.9—2014 National food safety standard Food microbiological examination:Examination of Campylobacter jejuni[S].Beijing:China Standards Press.2014. [8] 中华人民共和国国家卫生和计划生育委员会,国家食品药品监督管理总局.GB 4789.10—2016 食品安全国家标准 食品微生物学检验 金黄色葡萄球菌检验[S].北京:中国标准出版社,2016. National Health and Family Planning Commission of the People′s Republic of China,China Food and Drug Administration.GB 4789.10—2016 National food safety standard Food microbiological examination:Examination of Staphylococcus aureus[S].Beijing:China Standards Press,2016. [9] 中华人民共和国国家卫生和计划生育委员会.GB 4789.11—2014 食品安全国家标准 食品微生物学检验 β型溶血性链球菌检验[S].北京:中国标准出版社,2014. National Health and Family Planning Commission of the People′s Republic of China.GB 4789.11—2014 National food safety standard Food microbiological examination:Examination of Streptococcus[S].Beijing:China Standards Press,2014. [10] 中华人民共和国国家卫生和计划生育委员会,国家食品药品监督管理总局.GB 4789.12—2016 食品安全国家标准 食品微生物学检验 肉毒梭菌及肉毒毒素检验[S].北京:中国标准出版社,2016. National Health and Family Planning Commission of the People′s Republic of China,China Food and Drug Administration.GB 4789.12—2016 National food safety standard Food microbiological examination:Examination of Clostridiumbotulinum and otulinumtoxin[S].Beijing:China Standards Press,2016. [11] 中华人民共和国国家卫生部.GB 4789.13—2012 食品安全国家标准 食品微生物学检验 产气荚膜梭菌检验[S].北京:中国标准出版社,2012. Ministry of Health of the People′s Republic of China.GB 4789.13—2012 National food safety standard Food microbiological examination:Examination of Clostridiumperfringens[S].Beijing:China Standards Press,2012. [12] 中华人民共和国国家卫生和计划生育委员会.GB 4789.14—2014 食品安全国家标准 食品微生物学检验 蜡样芽胞杆菌检验[S].北京:中国标准出版社,2014. National Health and Family Planning Commission of the People′s Republic of China,China Food and Drug Administration.GB 4789.14—2014 National food safety standard Food microbiological examination:Examination of Bacillus cereus[S].Beijing:China Standards Press,2014. [13] 中华人民共和国国家卫生部.GB/T 4789.29—2003 食品卫生微生物学检验 椰毒假单胞菌酵米面亚种检验[S].北京:中国标准出版社,2003. Ministry of Health of the People′s Republic of China.GB/T 4789.29—2003 Microbiological examination of food hygiene-Examination of Pseudomonas cocovenenans subsf.farinofermentans[S].Beijing:China Standards Press,2003. [14] 中华人民共和国国家卫生部.GB 4789.38—2012 食品安全国家标准 食品微生物学检验 大肠埃希氏菌计数[S].北京:中国标准出版社,2012. Ministry of Health of the People′s Republic of China.GB 4789.38—2012 National food safety standard Food microbiological examination:Examination of Escherichiacoli[S].Beijing:China Standards Press,2012. [15] 中华人民共和国国家卫生和计划生育委员会,国家食品药品监督管理总局.GB 4789.30—2016 食品安全国家标准 食品微生物学检验 单核细胞增生李斯特氏菌检验[S].北京:中国标准出版社,2016. National Health and Family Planning Commission of the People′s Republic of China,China Food and Drug Administration.GB 4789.30—2016 National food safety standard Food microbiological examination:Examination of Listeria monocytogenes[S].Beijing:China Standards Press,2016. [16] 中华人民共和国国家卫生和计划生育委员会,国家食品药品监督管理总局.GB 4789.36—2016 食品安全国家标准 食品微生物学检验 大肠埃希氏菌O157:H7/NM检验[S].北京:中国标准出版社,2016. National Health and Family Planning Commission of the People′s Republic of China,China Food and Drug Administration.GB 4789.36—2016 National food safety standard Food microbiological examination:Examination of Escherichiacoli O157:H7/NM[S].Beijing:China Standards Press,2016. [17] 中华人民共和国国家卫生和计划生育委员会,国家食品药品监督管理总局.GB 4789.40—2016 食品安全国家标准 食品微生物学检验 克罗诺杆菌属(阪崎肠杆菌)检验[S].北京:中国标准出版社,2016. National Health and Family Planning Commission of the People′s Republic of China,China Food and Drug Administration.GB 4789.40—2016 National food safety standard Food microbiological examination:Examination of Cronobacter[S].Beijing:China Standards Press,2016. [18] 中华人民共和国国家卫生和计划生育委员会,国家食品药品监督管理总局.GB 4789.42—2016 食品安全国家标准 食品微生物学检验 诺如病毒检验[S].北京:中国标准出版社,2016. National Health and Family Planning Commission of the People′s Republic of China,China Food and Drug Administration.GB 4789.42—2016 National food safety standard Food microbiological examination:Examination of Norovirus[S].Beijing:China Standards Press,2016. [19] 中华人民共和国国家卫生和计划生育委员会.GB 4789.31—2013 食品安全国家标准 食品微生物学检验 沙门氏菌、志贺氏菌和致泻大肠埃希氏菌的肠杆菌科噬菌体诊断检验[S].北京:中国标准出版社,2013. National Health and Family Planning Commission of the People′s Republic of China.GB 4789.31—2013 National food safety standard Food microbiological examination:Bacteriophage diagnostic test for enterobacteriaceae for Salmonella,Shigella,and Escherichia coli causing[S].Beijing:China Standards Press,2013. [20] 中华人民共和国国家卫生和计划生育委员会,国家食品药品监督管理总局.GB 8538—2016 食品安全国家标准 饮用天然矿泉水检验方法[S].北京:中国标准出版社.2016. National Health and Family Planning Commission of the People′s Republic of China,China Food and Drug Administration.GB 8538—2016 National Food Safety Standard-Methods for examination of drinking natural mineral water[S].Beijing:China Standards Press.2016. [21] 中华人民共和国国家质量监督检验检疫总局.SN/T 3932—2014 出口食品中蜡样芽孢杆菌快速检测方法 实时荧光定量PCR法[S].北京:中国标准出版社,2014. State Administration of Quality Supervision,Inspection and Quarantine of the People′s Republic of China.SN/T 3932—2014 Rapid detection of Bacillus Cereus for export foods-Real-time PCR method[S].Beijing:China Standards Press,2014. [22] 中华人民共和国国家质量监督检验检疫总局.SN/T 2101—2016出口乳及乳制品中结核分枝杆菌检测方法 荧光定量PCR法[S].北京:中国标准出版社,2016. State Administration of Quality Supervision,Inspection and Quarantine of the People′s Republic of China.SN/T 2101—2016 Determination of Mycobacterium tuberculosis in milk and dairy products for export-Real-time PCR method[S].Beijing:China Standards Press,2016. [23] 中华人民共和国国家质量监督检验检疫总局.SN/T 2754.4—2011 出口食品中致病菌环介导恒温扩增(LAMP)检测方法 第4部分:单核细胞增生李斯特菌[S].北京:中国标准出版社,2011. State Administration of Quality Supervision,Inspection and Quarantine of the People′s Republic of China.SN/T 2754.4—2011 Loop-mediated isothermal amplification detection method for pathogens in export food-Part 4:Listeria monocytogenes[S].Beijing:China Standards Press,2011. [24] 河北省卫生和计划生育委员会.DB S13/008—2017 食品安全地方标准 肉及肉制品中沙门氏菌环介导等温扩增(LAMP)检测方法[S].石家庄:河北省卫生和计划生育委员会,2017. Health Commission of Hebei Province.DB S13/008—2017 Loop-mediated isothermal amplification detection method for Salmonella in meat and meat products[S].Shijiazhuang:Health and Family Planning Commission of Hebei,2017. [25] 吉林省市场监督管理厅.DB 22/T 2911—2018 贝类中副溶血性弧菌检测 微滴数字PCR法[S].长春:吉林省市场监督管理厅,2018. Jilin Market Supervision and Management Department.DB 22/T 2911—2018 Determination of Vibrio parahemolyticus in shellfish-droplet digital PCR method[S].Changchun:Jilin market supervision and management department,2018. [26] 中华人民共和国国家质量监督检验检疫总局.SN/T 3872—2014 出口食品中四种致病菌检测方法 MALDI-TOF-MS法[S].北京:中国标准出版社,2014. State Administration of Quality Supervision,Inspection and Quarantine of the People′s Republic of China.SN/T 3872—2014 Detection of four pathogens in food for export MALDI-TOF-MS method[S].Beijing:China Standards Press,2014. [27] 张飞燕,赵玲,金洁,等.多重 PCR技术在实验动物病原检测中的应用[J].中国比较医学杂志,2018,28(10):111-116. ZHANG F Y,ZHAO L,JIN J,et al.Application of multiplex PCR to detect experimental animal pathogens[J].Chinese Journal of Comparative Medicine,2018,28(10):111-116. [28] CHAMBERLAIN J S,GIBBS R A,RRANIER J E,et al.Deletion screeningof the Duchenne muscular dystrophy locus via multiplex DNA amplification[J].Nucleic Acids Res,1988,16(23):11 141-11 156. [29] 毕艳妮,宋宇,马淑青,等.多重PCR技术在病原微生物检测中的应用进展[J].中国微生态学杂志,2014,26(12):1 476-1 478. BI Y N,SONG Y,MA S Q,et al.Progress in the application of multiplex PCR in detection of pathogenic microorganisms[J].Chinese Journal of Microecology,2014,26(12):1 476-1 478. [30] CHEN Q,QIU M Y,FU Y F,et al.Establishment of detection method of suspension liquid chip of 4 kinds of respiratory viral[J].J Parasitic Biol,2013,8(2):97-101. [31] 李聪,刘健慧,李志辉.4种食源性致病菌的多重PCR快速检测方法研究[J].食品研究与开发,2019,40(21):164-169. LI C,LIU J H,LI Z H,et al.Study on multiplex PCR of rapid detection method for four foodborne pathogens[J].Food Research and Development,2019,40(21):164-169. [32] 杨鑫,王红宁,张安云,等.大肠杆菌氯霉素类耐药基因三重PCR检测试剂盒的研究与应用[J].中国兽医杂志,2009,45(2):11-13. YANG X,WANG H N,ZHANG A Y,et al.Development of multiplex PCR detecting kit and its applicaton to the detection of chloramphenicols resistant genes in Escherichia coli[J].Chinese Journal of Veterinary Medicine,2009,45(2):11-13. [33] LIM Y H,HIROSE K,IZUMIYA H,et al.Multiplex polymerase chain reaction assay for selective detection of Salmonella enterica serovar typhimurium[J].Jpn J Infect Dis,2003,56(4):151-155. [34] 李伟杰,于建慧,魏财文,等.产气荚膜梭菌毒素型菌落多重PCR方法的建立及应用[J].西北农业学报,2016,25(6):823-827. LI W J,YU J H,WEI C W,et al.Establishment and application of colony multiplex pcr assay for toxinotyping Ciostridium perfringens[J].Acta Agriculturae Boreali-occidentalis Sinica,2016,25(6):823-827. [35] 王倩,徐新明,胡蕊,等.福氏志贺氏菌QRDR基因gyrA,parC及毒力基因ipaH的多重PCR检测[J].临床与病理杂志,2019,39(1):22-26. WANG Q,XU X M,HU R,et al.Multiplex PCR detection of QRDR genes gyrA,parC and virulence gene ipaH of Shigella flexneri[J].Journal of International Pathology and Clinical Medicine,2019,39(1):22-26. [36] 楼秀芹,斯国静,戚建江,等.食品中阪崎肠杆菌多重PCR检测方法的建立[J].中国卫生检验杂志,2014,24(2):159-163. LOU X Q,SI G J,QI J J,et al.Detection of Enterobacter sakazakii in food by multi - plex PCR[J].Chinses Journal of Health Laboratory Technology,2014,24(2):159-163. [37] 黎明,阮佳,李永新.食源性阪崎肠杆菌的双重PCR检测方法研究[J].中国卫生检验杂志,2013,23(6):1 473-1 476. LI M,RUAN J,LI Y X,et al.Detection of enterobacter sakazakii in food by duplex PCR[J].Chinses Journal of Health Laboratory Technology,2013,23(6):1 473-1 476. [38] 陈伟.六种食源性致病微生物PCR检测及固相化试剂盒的研究[D].重庆:重庆大学,2008. CHEN W.Study on the detection of six food-borne bacterial pathogens by PCR and multiplex PCR detection kit[D].Chongqing:Chongqing University,2008. [39] 郭倩倩.多重PCR技术和重组DNA技术在水产品安全中的应用[D].上海:上海海洋大学,2010. GUO Q Q.Application of multiplex PCR and recombinant DNA technologies in the control of aquatic product security[D].Shanghai:Shanghai Ocean University,2010. [40] 周杨,万强,蔡芷荷,等.食品中克罗诺杆菌属双重PCR检测试剂盒的评价[J].微生物学通报,2019,46(9):2 282-2 291. ZHOU Y,WAN Q,CAI Z H,et al.Evaluation of duplex PCR detection kit for detection of Cronobacter in food samples[J].Microbiology China,2019,46(9):2 282-2 291. [41] 刘文干.11种肠道致病菌核酸多重PCR检测试剂盒及其应用:中国,CN105525031A[P].2016-04-27. LIU W G.Multiple PCR kits for detection of 11 intestinal pathogenic bacteria nucleic acids and their applications:China,CN105525031A[P].2016-04-27. [42] 王雷.十四种食源性致病菌多重PCR检测引物组和试剂盒:中国,CN103484546B[P].2015-07-01. WANG L.Primer sets and kits for multiple PCR detection of 14 foodborne pathogenic bacteria,China:CN103484546B[P].2015-07-01. [43] 宋丽萍,姜洁,李玮,等.食源性致病菌快速检测技术研究进展[J].食品安全质量检测学报,2015,6(9):3 441-3 446. SONG L P,JIANG J,LI W,et al.Research progress on rapid detection of foodborne bacterial pathogens[J].Journal of Food Safety and Quality,2015,6(9):3 441-3 446. [44] OWAIS M,KAEMIS,TUFAIL S.An alternative chemical redox method for the production of bispecific antibodies:Implication in rapid detection of food borne pathogens[J].PLoS One,2014,9(3):e91 255.
2021, Vol. 47 
