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2/2018 vol. 68
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Rapid and Specific Detection of Escherichia coli O157:H7 in Ground Beef Using Immunomagnetic Separation Combined with Loop-Mediated Isothermal Amplification
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Publication date: 2018-06-30
 
Pol. J. Food Nutr. Sci. 2018;68(2):115–123
DOI: https://doi.org/10.1515/pjfns-2017-0014
 
 
Article (PDF)
References (48) Citations (5)
 
KEYWORDS
immunomagnetic separation
LAMP
Escherichia coli
EHEC O157:H7
rfbE gene
ground beef
 
ABSTRACT
Escherichia coli O157:H7 is well known for many foodborne outbreaks that lead to fatal infections in human being worldwide. The objective of this study was to develop a rapid and sensitive method for detection of EHEC O157:H7 from ground beef using a method that combined immunomagnetic separation (IMS) with loop-mediated isothermal amplification (LAMP). The EHEC O157:H7 cells were separated with Dynabeads coated with anti-EHEC O157:H7 after a short enrichment for 4 h. Then, EHEC O157:H7 was identified by LAMP assay for amplifying and detecting the rfbE gene, which is highly conserved in all EHEC O157:H7 strains and exhibits strain-specific gene expression. The LAMP method results analyzed with real time turbidity measurements showed a high specificity and sensitivity, with a positive detection rate of amplification of EHEC O157:H7 DNA diluted to a minimum equivalent concentration of 1.8 × 101 CFU/mL, which was 10 times more sensitive than the conventional PCR assay. The IMS followed with LAMP could capture and detect a bacterial concentration as low as 3×101 CFU/mL from the meat samples, which was close to the sensitivity of LAMP assay with pure culture. IMS combined with realistic LAMP method is a simple, rapid, highly specific gene amplification technology that is suitable for implementing as a screening assay in basic laboratory and field test for detecting food contamination.
 
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An evaluation of immunomagnetic separation-real-time PCR (IMS-RTiPCR) combined assay for rapid and specific detection of Escherichia coli O157:H7 in raw milk and ground beef
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Food Science and Technology
 
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Immunomagnetic separation: An effective pretreatment technology for isolation and enrichment in food microorganisms detection
Zhouli Wang, Rui Cai, Zhenpeng Gao, Yahong Yuan, Tianli Yue
Comprehensive Reviews in Food Science and Food Safety
 
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Newly isolated Coliphages for Bio-controlling Multidrug Resistant Escherichia coli strains
Mostafa Elbahnasawy, ElSayed ElSayed, Mohamed Azzam
Environmental Nanotechnology, Monitoring & Management
 
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Rapid on-site detection of viable Escherichia coli O157: H7 in lettuce using immunomagnetic separation combined with PMAxx-LAMP and nucleic acid lateral flow strip
Yuanyi Wen, Youjiang Tan, Lichao Zhao, Xinrui Lv, Li Lin, Dezhi Liang, Li Wang
Microchemical Journal
 
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Review in isothermal amplification technology in food microbiological detection
Xiang Li, Xiaolong Zhang, Xiaoli Shi, Hongfei Shi, Zhouping Wang, Chifang Peng
Food Science and Biotechnology
 
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