Rapid and specific detection of Escherichia coli O157:H7 in ground beef using immunomagnetic separation combined with loop-mediated isothermal amplification

• Autorzy: Qin Y. , Puthiyakunnon S. , Zhang Y. , Wu X. , Boddu S. , Luo B. , Fan H.
• Języki publikacji: EN

Abstrakty

EN

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 amplifi cation (LAMP). The EHEC O157:H7 cells were separated with Dynabeads coated with antiEHEC O157:H7 after a short enrichment for 4 h. Then, EHEC O157:H7 was identifi ed 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 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.

Słowa kluczowe

EN

human infection; Escherichia coli; food-borne disease; rapid detection; specific detection; ground beef; beef; immunomagnetic separation; loop-mediated isothermal amplification; rfbE gene

• Wydawca: -
• Czasopismo: Polish Journal of Food and Nutrition Sciences
• Rocznik: 2018
• Tom: 68
• Numer: 2
• Opis fizyczny: p.115-123,fig.,ref.

Twórcy

autor

Qin Y.

• Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, 510515, China

autor

Puthiyakunnon S.

• Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, 510515, China

autor

Zhang Y.

• Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, 510515, China

autor

Wu X.

• Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, 510515, China

autor

Boddu S.

• Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, 510515, China

autor

Luo B.

• Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, 510515, China

autor

Fan H.

• Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, 510515, China

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Identyfikatory

DOI

10.1515/pjfns-2017-0014