Rapid and Specific Detection of Escherichia coli O157:H7 in Ground Beef Using Immunomagnetic Separation Combined with Loop-Mediated Isothermal Amplification
Publication date: 2018-06-30
Pol. J. Food Nutr. Sci. 2018;68(2):115–123
KEYWORDS
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.
CITATIONS (2):
1.
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
TABAN MERCANOGLU, Sait AYTAC
Food Science and Technology
TABAN MERCANOGLU, Sait AYTAC
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
Zhouli Wang, Rui Cai, Zhenpeng Gao, Yahong Yuan, Tianli Yue
Comprehensive Reviews in Food Science and Food Safety
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