期刊
FOOD CONTROL
卷 157, 期 -, 页码 -出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.foodcont.2023.110171
关键词
Real-time PCR; O-serotypes; Escherichia coli; Serogrouping; Multiplex PCR; Foodborne outbreaks
A novel multiplex real-time PCR method was developed for rapid and accurate serogrouping of Escherichia coli O-serotypes, enabling identification in food environments.
Escherichia coli O-serotypes are associated with pathogenesis; thus, rapid and accurate serogrouping is required to prevent foodborne outbreaks. In this study, a novel multiplex real-time PCR method was developed for E. coli serogrouping to overcome some limitations of conventional and in silico serogrouping. Two O-serotype genes, wzy and wzx, were selected, and 21 primer-probe sets were designed for determining O-serotypes. To evaluate this, 11 O-serotype reference strains and 43 environmental isolates were tested: serogrouping took <40 min and the detection limit was 1-10 pg DNA, indicating that rapid identification was achieved with high sensitivity and without false-positive results. Subsequent food tests revealed that the method worked perfectly even at 10(1) CFU per ground beef sample; thus, O-serotypes were identifiable in food environments. The developed multiplex real-time PCR serogrouping method can rapidly, inexpensively, and accurately identify E. coli O-serotypes; therefore, it could help prevent foodborne outbreaks.
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