Applicability of a multiplex PCR to detect the seven major Shiga toxin-producing Escherichia coli based on genes that code for serogroup-specific O-antigens and major virulence factors in cattle feces

Foodborne Pathogens and Disease
Jianfa BaiT G Nagaraja

Abstract

An 11-gene multiplex polymerase chain reaction (mPCR) was developed based on genes that code for serogroup-specific O-antigens and four major virulence factors (intimin, enterohemorrhagic hemolysin, and Shiga toxins [Stx] 1 and 2), to detect O157 and the "top six" non-O157 (O26, O45, O103, O111, O121, and O145) Shiga toxin-producing Escherichia coli (STEC). The assay specificity was validated with pure cultures of seven major STEC (185 strains), 26 other STEC (65 strains), non-STEC (five strains), and 33 strains of other genera and species. Sensitivity of the assay with cattle fecal sample spiked with pooled cultures of seven major STEC was 10⁵ colony-forming units (CFU)/g before enrichment and 10² CFU/g after enrichment. The applicability of the assay to detect STEC in fecal samples (n=50), before and after enrichment, was evaluated by comparing with culture-based methods for O26, O111, and O157. The mPCR assay of 50 fecal samples showed seven (14%) positive before enrichment and 23 (46%) positive after enrichment for one or more of the seven O-groups. Overall, 17 isolates from 17 fecal samples and 27 isolates (four for O26, three for O45, and 20 for O103) from 19 fecal samples were obtained, by culture-based methods, for O157...Continue Reading

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Citations

Mar 6, 2013·Foodborne Pathogens and Disease·David A DargatzGary A Anderson
Jun 21, 2015·Journal of Microbiological Methods·Loreen R StrombergHarshini Mukundan
Apr 22, 2015·International Journal of Environmental Research and Public Health·Benson Chuks IwerieborAnthony I Okoh
Mar 25, 2015·Journal of Applied Microbiology·J E WellsW T Oliver
May 18, 2016·Foodborne Pathogens and Disease·Faruk PehlivanogluDilek Ozturk
May 12, 2016·Journal of Medical Microbiology·Benson C IwerieborAnthony I Okoh
Sep 16, 2016·Frontiers in Cellular and Infection Microbiology·Travis A WoodsAlina Deshpande
Mar 5, 2016·Journal of Food Protection·Zachary R StrombergRodney A Moxley
Mar 21, 2013·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Kudakwashe MagwedereChitrita DeBroy
Feb 10, 2018·Current Microbiology·Gentry L LewisRodney A Moxley
Jul 24, 2018·Foodborne Pathogens and Disease·Elis Caroline Celestina Dos SantosEduardo Eustáquio de Souza Figueiredo
Jul 25, 2018·Microorganisms·Kim StanfordTim A McAllister
Jan 23, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Maria Grazia CusimanoMaria Vitale

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