A Modified Molecular Beacons-Based Multiplex Real-Time PCR Assay for Simultaneous Detection of Eight Foodborne Pathogens in a Single Reaction and Its Application

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	A Modified Molecular Beacons-Based Multiplex Real-Time PCR Assay for Simultaneous Detection of Eight Foodborne Pathogens in a Single Reaction and Its Application
	Hu, Qinghua ; Lyu, DongYue ; Shi, Xiaolu ; Jiang, Yixiang ; Lin, Yiman ; Li, Yinghui ; Qiu, Yaqun ; He, Lianhua ; Zhang, Ran ; Li, Qingge ; Li QG(李庆阁)
刊名	http://dx.doi.org/10.1089/fpd.2013.1607
	2014-03-01
关键词	POLYMERASE-CHAIN-REACTION SPECIES-SPECIFIC IDENTIFICATION ESCHERICHIA-COLI LISTERIA-MONOCYTOGENES YERSINIA-ENTEROCOLITICA CAMPYLOBACTER-JEJUNI SALMONELLA SPP. SHIGELLA TECHNOLOGY INFECTIONS
英文摘要	National Natural Science Foundation of China [81071433]; Guangdong Provincial Natural Science Foundation [8151802003000006]; Basic Research Project of Shenzhen [JC201104220329A]; Foodborne disease outbreaks are often caused by one of the major pathogens. Early identification of the causal pathogen is crucial for disease control and prevention. We describe a real-time polymerase chain reaction (rtPCR) assay that can identify, in a single reaction, up to eight common foodborne bacterial pathogens, including Salmonella enterica subsp. enterica, Listeria monocytogenes, Escherichia coli O157, Vibrio parahaemolyticus, V. vulnificus, Campylobacter jejuni, Enterobacter sakazakii, and Shigella spp. This multiplex rtPCR assay takes advantage of modified molecular beacons and the multicolor combinational probe coding strategy to discriminate each pathogen and the homo-tag assisted non-dimer (HAND) system to prevent dimer formation. The detection limits of the assay ranged from 1.3x10(3) colony-forming units (CFU)/g stool (L. monocytogenes) to 1.6x10(4) CFU/g stool (Shigella spp.). The target genes were 100% specific as assessed on 986 reference strains covering 41 species since no cross-reactions were observed. The assay was applied to the detection of foodborne pathogens in 11,167 clinical samples and the results were compared with culture methods for further validation. The sensitivity and specificity of the rtPCR were 100% and 99%, respectively. When performed in a 96-well rtPCR system, more than 90 samples could be analyzed within 3 h. Given the high accuracy, sensitivity, specificity, and short turn-around time, the established assay could be used for the rapid and reliable identification of the causative pathogens responsible for a certain foodborne disease outbreak and rapid screening of these major foodborne pathogens in laboratory-based surveillance of outpatient clinical samples or even food samples.
语种	英语
出版者	MARY ANN LIEBERT, INC
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/90824]
专题	生命科学－已发表论文
推荐引用方式 GB/T 7714	Hu, Qinghua,Lyu, DongYue,Shi, Xiaolu,et al. A Modified Molecular Beacons-Based Multiplex Real-Time PCR Assay for Simultaneous Detection of Eight Foodborne Pathogens in a Single Reaction and Its Application[J]. http://dx.doi.org/10.1089/fpd.2013.1607,2014.
APA	Hu, Qinghua.,Lyu, DongYue.,Shi, Xiaolu.,Jiang, Yixiang.,Lin, Yiman.,...&李庆阁.(2014).A Modified Molecular Beacons-Based Multiplex Real-Time PCR Assay for Simultaneous Detection of Eight Foodborne Pathogens in a Single Reaction and Its Application.http://dx.doi.org/10.1089/fpd.2013.1607.
MLA	Hu, Qinghua,et al."A Modified Molecular Beacons-Based Multiplex Real-Time PCR Assay for Simultaneous Detection of Eight Foodborne Pathogens in a Single Reaction and Its Application".http://dx.doi.org/10.1089/fpd.2013.1607 (2014).