Rapid and Specific Detection of Escherichia coli O157:H7 in Ground Beef Using Immunomagnetic Separation Combined with Loop-Mediated Isothermal Amplification
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 (I...
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| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences
2018-06-01
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| Series: | Polish Journal of Food and Nutrition Sciences |
| Subjects: | |
| Online Access: | http://www.degruyter.com/view/j/pjfns.2018.68.issue-2/pjfns-2017-0014/pjfns-2017-0014.xml?format=INT |
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| Summary: | 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-specifi c gene expression. The LAMP method results analyzed with real time turbidity measurements showed a high specificity and sensitivity, with a positive detection rate of amplifi cation 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 amplifi cation technology that is suitable for implementing as a screening assay in basic laboratory and field test for detecting food contamination. |
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| ISSN: | 2083-6007 |