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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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 |
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| 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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| author | Qin Yu Puthiyakunnon Santhosh Zhang Yiduo Wu Xianbo Boddu Swapna Luo Binde Fan Hongying |
| author_facet | Qin Yu Puthiyakunnon Santhosh Zhang Yiduo Wu Xianbo Boddu Swapna Luo Binde Fan Hongying |
| author_sort | Qin Yu |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-955cfc0a9d6a4c0baa2b1bbe98437c37 |
| institution | Kabale University |
| issn | 2083-6007 |
| language | English |
| publishDate | 2018-06-01 |
| publisher | Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences |
| record_format | Article |
| series | Polish Journal of Food and Nutrition Sciences |
| spelling | doaj-art-955cfc0a9d6a4c0baa2b1bbe98437c372025-02-02T00:52:05ZengInstitute of Animal Reproduction and Food Research of the Polish Academy of SciencesPolish Journal of Food and Nutrition Sciences2083-60072018-06-0168211512310.1515/pjfns-2017-0014pjfns-2017-0014Rapid and Specific Detection of Escherichia coli O157:H7 in Ground Beef Using Immunomagnetic Separation Combined with Loop-Mediated Isothermal AmplificationQin Yu0Puthiyakunnon Santhosh1Zhang Yiduo2Wu Xianbo3Boddu Swapna4Luo Binde5Fan Hongying6Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, 510515, ChinaGuangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, 510515, ChinaGuangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, 510515, ChinaGuangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, 510515, ChinaGuangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, 510515, ChinaGuangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, 510515, ChinaGuangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, 510515, ChinaEscherichia 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.http://www.degruyter.com/view/j/pjfns.2018.68.issue-2/pjfns-2017-0014/pjfns-2017-0014.xml?format=INTimmunomagnetic separationLAMPEscherichia coliEHEC O157:H7rfbE geneground beef |
| spellingShingle | Qin Yu Puthiyakunnon Santhosh Zhang Yiduo Wu Xianbo Boddu Swapna Luo Binde Fan Hongying Rapid and Specific Detection of Escherichia coli O157:H7 in Ground Beef Using Immunomagnetic Separation Combined with Loop-Mediated Isothermal Amplification Polish Journal of Food and Nutrition Sciences immunomagnetic separation LAMP Escherichia coli EHEC O157:H7 rfbE gene ground beef |
| title | Rapid and Specific Detection of Escherichia coli O157:H7 in Ground Beef Using Immunomagnetic Separation Combined with Loop-Mediated Isothermal Amplification |
| title_full | Rapid and Specific Detection of Escherichia coli O157:H7 in Ground Beef Using Immunomagnetic Separation Combined with Loop-Mediated Isothermal Amplification |
| title_fullStr | Rapid and Specific Detection of Escherichia coli O157:H7 in Ground Beef Using Immunomagnetic Separation Combined with Loop-Mediated Isothermal Amplification |
| title_full_unstemmed | Rapid and Specific Detection of Escherichia coli O157:H7 in Ground Beef Using Immunomagnetic Separation Combined with Loop-Mediated Isothermal Amplification |
| title_short | Rapid and Specific Detection of Escherichia coli O157:H7 in Ground Beef Using Immunomagnetic Separation Combined with Loop-Mediated Isothermal Amplification |
| title_sort | rapid and specific detection of escherichia coli o157 h7 in ground beef using immunomagnetic separation combined with loop mediated isothermal amplification |
| topic | immunomagnetic separation LAMP Escherichia coli EHEC O157:H7 rfbE gene ground beef |
| url | http://www.degruyter.com/view/j/pjfns.2018.68.issue-2/pjfns-2017-0014/pjfns-2017-0014.xml?format=INT |
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