Microsoft Word 杨纯佳二校% 出4726、4730

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1 7 12 Vol. 7 No Journal of Food Safety and Quality Dec., 2016 PCR 杨纯佳, 张 * 娟, 周臣清, 霍丽斯, 崔海萍, 王力清 (, ( ), ) 摘要 : PCR, PCR,, PCR PCR PCR PCR O157:H7 PCR, 关键词 : PCR; PCR; PCR; Research progress on the application of digital PCR in detection of foodborne pathogenic microorganisms YANG Chun-Jia, ZHANG Juan, ZHOU Chen-Qing, HUO Li-Si, CUI Hai-Ping, WANG Li-Qing * (Guangdong Testing Institute of Product Quality Supervision, China National Quality Supervision and Testing Center for Foods (Guangdong), Foshan , China) ABSTRACT: Foodborne diseases are mostly caused by foodborne pathogens, so it is very important to establish effective detection methods of foodborne pathogenic microorganisms for food safety risk monitoring and people's health ensuring. Compared with the traditional PCR, digital PCR has good accuracy and reproducibility, which has provided a new platform for rapid detection of food safety. This article mainly summarized the basic principles, varieties, application and research progress of digital PCR technology including droplet digital PCR and chip digital PCR. The applications of digital PCR technology in detection of Salmonella, Escherichia coli O157:H7, Listeria monocytogenes, Enterobacter cloacae, Staphylococcus aureus and other foodborne pathogenic microorganisms were also discussed in depth. The technology digital PCR has gotten good applications in quantitative detection of genetically modified organisms and animal derived ingredients, but was remained to be better in the application of foodborne pathogenic microorganisms. KEY WORDS: digital PCR; droplet digital PCR; chip digital PCR; foodborne pathogenic microorganisms 基金项目 : (2016A ) Fund: Supported by Science and Technology Project of Guangdong Province (2016A ) * 通讯作者 :,, sdwlq@21cn.com *Corresponding author: WANG Li-Qing, Professor Senior Engineer, Guangdong Testing Institute of Product Quality Supervision, Foshan , China sdwlq@21cn.com

2 12, : PCR 引言,,,,, 70% [1] [2],, [3], PCR, [4-6], PCR,,,, DNA,, DNA, [7] PCR PCR PCR PCR PCR PCR 2 数字 PCR 技术概述 PCR(digital PCR, dpcr) 1992, Sykes [8] PCR, PCR PCR, PCR C t ( ),,,, [9], PCR PCR, PCR PCR [10-14] PCR dpcr(droplet dpcr, ddpcr) dpcr(chip dpcr, cdpcr) PCR PCR,,, PCR,, 1, 0, [15,16] ddpcr PCR(qPCR),,, [17-20] cdpcr PCR,, [21] [22] PCR, PCR 3 dpcr 在食源性致病微生物检测中的应用 3.1 dpcr 检测食品中沙门氏菌,,, [23,24] [25] SD-PMA-ddPCR( - - PCR, sodium deoxycholate-propidium monoazide-droplet dpcr), ddpcr 8.0 copy/20 μl, 10 2 CFU/mL SD-PMA-ddPCR, Rothrock [26] ddpcr,, ddpcr dpcr, 3.2 ddpcr 定量检测大肠杆菌 O157: H7 (enterohemorrhagic Escherichia coli, EHEC) O157:H7,,,, [27] O157:H7, 1982 O157:H7 O157:H7 [28-30] 1999 O157:H7,

3 , 177, 7 [27], [31] O157: H7 rfbe, ddpcr ddpcr 300 nmol/l O157:H7 DNA 4~ copy/20 μl ddpcr, ddpcr DNA 760~88400 copy/20 μl, (RSD<5%) 4 copy/20 μl, 3 copy/20 μl, ddpcr, 13, PCR Bian [32] ddpcr O157:H7, PCR, ddpcr 10 CFU/mL PCR, dpcr 3.3 dpcr 定量检测单核细胞增生李斯特氏菌 (Listeria monocytogenes),,,, 20%~50% [33] 4,,, [34] [35] SD-PMA-ddPCR, (sodium deoxycholate, SD), (propidium monoazide, PMA) DNA, DNA ddpcr SD-PMA-ddPCR 2.0 copy/20 μl, Rothrock [26] ddpcr,, ddpcr Bian [32] ddpcr, 10 CFU/mL SD-PMA-ddPCR, 3.4 dpcr 定量检测阴沟肠杆菌 1980 (Enterobacter sakazakii) ( ), 50% [36] dpcr, qpcr,, 8.9 copy/μl,, 3.5 dpcr 定量检测金黄色葡萄球菌 (Staphylococcus aureus, SA),, [37-39], 33%, 45%, [40-42] Kelley [43] ddpcr, qpcr ddpcr SA0140(SA) (MECA) 397, qpcr ddpcr, ddpcr qpcr,,, dpcr, 4 结语与展望 dpcr,, :(1)dPCR, ; (2) PCR, ;(3)dPCR PCR, PCR PCR,,, ;, 参考文献 [1] Groundwater PW, Todd A, Worsley A J, et al. The technology and techniques used in the detection of pathogenic bacteria [J]. Pharm J, 2009, 283(7569): [2]. PCR [J]., 2006, CFI(11):

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Microsoft Word %

Microsoft Word % 8 1 Vol. 8 No. 1 2017 1 Journal of Food Safety and Quality Jan., 2017 2015 孙寒松 *, 王德宇 (, 132001) 摘要 : 目的, 方法 2014 结果 2015 10 240, 19, 7.92%, 7, 8, 3, 1 ; 3 : 57.14%, 50%, 2%; 结论,, 关键词 : ; ; Analysis of

Microsoft Word 杨纯佳 二校% 出4726、4730

Microsoft Word 杨纯佳二校% 出4726、4730