Food Hygiene and Safety Science (Shokuhin Eiseigaku Zasshi)
Online ISSN : 1882-1006
Print ISSN : 0015-6426
ISSN-L : 0015-6426
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DNA Extraction for Sensitive Detection of Shiga Toxin-Producing Escherichia coli in Food by Real-Time PCR Assays
Tetsuya MoriSayaka Nagao-SatoKanae KishinoToyohiko NambaYukiko Hara-Kudo
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2019 Volume 60 Issue 6 Pages 183-186

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Abstract

Alkali-heat DNA extraction, a rapid and economical method, was evaluated for use in the detection of Shiga toxin-producing Escherichia coli in food using real-time PCR assays. Alkali-heat DNA extracts led to highly sensitive detection (102–104 CFU/mL) of stx and O-antigen genes in beef liver, ground beef, sliced pork, cheese, lettuce, radish sprouts, tomato, and spinach, equivalent to the sensitivity obtained using a commercial DNA extraction kit that utilizes proteinase K lysis, and silica membrane purification. Although there were differences in DNA concentration and purity between DNA extraction methods, the sensitivity of real-time PCR assays was similar. These results indicate that alkali-heat DNA extraction is a viable method when testing food products with real-time PCR assays for the presence of stx and O-antigen genes.

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© 2019 Japanese Society for Food Hygiene and Safety
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