Universal Primer-Multiplex PCR Approach for Simultaneous Detection of Escherichia coli, Listeria monocytogenes, and Salmonella spp. in Food Samples

被引:38
|
作者
Yuan, Yanfang [1 ]
Xu, Wentao [1 ,2 ]
Zhai, Zhifang [1 ]
Shi, Hui [1 ]
Luo, Yunbo [1 ]
Chen, Zhuojun [1 ]
Huang, Kunlun [1 ,2 ]
机构
[1] China Agr Univ, Coll Food Sci & Nutr Engn, Beijing 100083, Peoples R China
[2] Minist Agr, Supervis Inspect & Testing Ctr Genetically Modifi, Beijing 100083, Peoples R China
关键词
Escherichia coli; Listeria monocytogenes; Salmonella spp; simultaneous detection; universal primer-multiplex PCR; REAL-TIME PCR; POLYMERASE-CHAIN-REACTION; MULTISTATE OUTBREAK; RAPID DETECTION; O157-H7; INFECTION; ASSAYS; IDENTIFICATION; PATHOGENS; PRODUCTS;
D O I
10.1111/j.1750-3841.2009.01321.x
中图分类号
TS2 [食品工业];
学科分类号
0832 ;
摘要
Escherichia coli, Listeria monocytogenes, and Salmonella spp. are 3 kinds of the most important food-borne human pathogens. Traditional microbiological analysis is labor-intensive, time-consuming, and easily contaminated, thus producing false positive signals; it also involves much subjectivity judgments. Multiplex-PCR could be applied to detect multiple target organisms simultaneously to save time and labor, but there is always disproportionate amplification resulting from the disparity of different primers. To gain a rapid and sensitive method, a universal primer-multiplex PCR system (UP-M-PCR) was developed and applied for simultaneous detection of the 3 organisms. This method simplified traditional multiplex-PCR reaction system and overcame its amplification disparities among different primers; moreover, it got a high specificity and sensitivity (85, 155, and 104 copies/reaction for E. coli O157, L. monocytogenes, and Salmonella spp., respectively). Compared with the time-consuming and laborious microbiological analysis, UP-M-PCR had a lower risk of cross-contamination without inoculation and incubation. Test results for 36 food samples showed that UP-M-PCR method got a relative accuracy of 91.77% when compared with traditional microbiological analysis. It could serve as a rapid screening method for pathogen detection and could detect target genes even in dead pathogenic cells. In addition, it has the potential to be performed in an automation mode and might find broader application in simultaneous detection of other multiple pathogens.
引用
收藏
页码:M446 / M452
页数:7
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