| Autor: |
Yu Qin, Puthiyakunnon, Santhosh, Yiduo Zhang, Xianbo Wu, Boddu, Swapna, Luo, Binde, Hongying Fan |
| Předmět: |
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| Zdroj: |
Polish Journal of Food & Nutrition Sciences; 2018, Vol. 68 Issue 2, p115-123, 9p, 1 Diagram, 2 Charts, 5 Graphs |
| Abstrakt: |
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. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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