Applicability of a multiplex PCR to detect O26, O45, O103, O111, O121, O145, and O157 serogroups of Escherichia coli in cattle feces1

Paddock, Zachary Dean; Shi, Xiaorong; Bai, Jianfa; Nagaraja, T. G.

doi:10.1016/j.vetmic.2011.11.017

Cited by 73 publications

(56 citation statements)

References 38 publications

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“…A large number of samples positive for O serogroups, by both culture-based and direct PCR methods, did not possess either Shiga toxin gene, indicating that cattle carry Shiga toxin gene-negative E. coli belonging to these seven O serogroups. This phenomenon has been previously observed with O157 (Bielaszewska et al, 2007;Wetzel and LeJeune, 2007;Cernicchiaro et al, 2009), and non-O157 STEC strains in cattle feces Paddock et al, 2012).…”

Section: Discussionsupporting

confidence: 81%

“…The detection of other STEC serogroups is likely due to nonspecific binding on the beads and subsequent use of a nondifferential medium. Isolation of STEC obtained by culture-based methods followed by PCR confirmation of serogroup and virulence genes, or the detection of gene markers using PCR directly on enriched samples, was shown to be advantageous for testing fecal samples that may harbor more than one serogroup and may not possess Shiga toxin or intimin genes Paddock et al, 2012). Other fecal prevalence studies (Cobbold et al, 2004;Renter et al, 2007) followed an FSIS-type approach (FSIS, 2010) by prescreening samples for Shiga toxin genes before applying cultural methods.…”

Section: Discussionmentioning

confidence: 99%

“…This issue should be further investigated in order to characterize STEC risk associated with cattle production systems. Paddock et al (2012) and Bai et al (2012) concluded that multiplex PCR for STEC serogroups is most applicable for confirming putative isolates. Our results support their findings that culture-based methods using specific IMS beads followed by PCR confirmation of serogroup and virulence genes identifies a greater number of O-positive samples than using PCR directly.…”

Section: Discussionmentioning

confidence: 99%

“…It may be that PCR detected genes carried by bacteria other than the targeted STEC serogroups. Methods for isolation and detection of non-O157 STEC in feces are still under development (Possé et al, 2008;Bai et al, 2012;Hedge et al, 2012;Paddock et al, 2012;Wylie et al, 2013); specific potential needs are to develop a selective medium to phenotypically identify STEC serogroups, improve specificity of IMS beads, validate molecular techniques for faster and more sensitive detection, and standardize protocols to accurately identify STEC.…”

Section: Discussionmentioning

confidence: 99%

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Prevalence of Shiga Toxin–Producing Escherichia coli and Associated Virulence Genes in Feces of Commercial Feedlot Cattle

Cernicchiaro

Cull

Paddock

et al. 2013

Foodborne Pathogens and Disease

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The objective of this study was to determine the prevalence of Shiga toxin-producing Escherichia coli (STEC) serogroups and associated virulence genes in feces of commercial feedlot cattle. During March to May 2011, fecal samples were collected from individual cattle (n = 960) in 10 cohorts (cattle subpopulations within a feedlot) comprising 17,148 total steers that originated from 48 backgrounding operations in six U.S. states. Fecal samples were enriched in E. coli broth and subjected to two detection protocols: (1) an 11-gene multiplex polymerase chain reaction (PCR) that identifies seven O serogroups (O26, O45, O103, O111, O121, O145, and O157) and four virulence genes (stx1, stx2, eae, and ehxA) applied to extracted total DNA (''direct PCR''); and (2) cultural procedures that involve immunomagnetic separation (IMS) with O26, O103, and O111 beads, plating on a nondifferential MacConkey agar, followed by the multiplex PCR of pooled colonies (''culture-based method''). Generalized linear mixed models were used to adjust prevalence estimates for clustering. Based on direct PCR detection, O157 (49.9%) was the most prevalent O serogroup followed by O26 (20.3%), O103 (11.8%), O121 (10.7%), O45 (10.4%), O145 (2.8%), and O111 (0.8%). Cumulative adjusted prevalence estimates were 22.3, 24.6, and 0.01% for O26, O103, and O111 serogroups, respectively, based on culture-based methods. However, prevalence varied significantly by cohort ( p-values < 0.05) for O26, O121, and O157 based on direct PCR, and for O26, O103, and O111 serogroups based on culture-based methods. Results of this study indicate that all seven STEC serogroups were identified in feedlot cattle feces, with O157, O26, and O103 being the most prevalent serogroups. A substantial proportion of serogroup-positive samples did not harbor Shiga toxin genes; thus, additional elucidation of the potential human health risk is required. Further evaluation of diagnostic methods for non-O157 STEC is needed given their impact on prevalence estimation.

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Section: Discussionsupporting

confidence: 81%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Prevalence of Shiga Toxin–Producing Escherichia coli and Associated Virulence Genes in Feces of Commercial Feedlot Cattle

Cernicchiaro

Cull

Paddock

et al. 2013

Foodborne Pathogens and Disease

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“…Os autores também identificaram o sorotipo O26 em 0,2% das amostras de pele, 1,5% das de fezes e em nenhuma carcaça, diferenciando-se deste estudo, que mostrou a presença da bactéria em 6,0% das amostras de pele, em nenhuma amostra de fezes e 2,0% das amostras de músculo externo da carcaça. Paddock et al (2012), verificando a aplicabilidade da PCR multiplex para detectar E. coli não-O157 e O157 em bovinos, identificaram a E. coli O157 em 49,0% e O26 em 82,0% das amostras, valores maiores que os obtidos neste trabalho, de 12,0% e 8,0%. Já Buvens et al (2012), ao estudarem STEC em casos de infecções na Bélgica, relataram menor ocorrên-cia de E. coli O26 (0,14%) em relação a este estudo (8,0%).…”

Section: Resultados E Discussão Ocorrência De Escherichia Coliunclassified

Ocorrência de Escherichia coli O157:H7 e O26 sorbitol negativas em matadouro frigorífico de bovino e suscetibilidade a antimicrobianos

et al. 2014

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Cattle beef could be a way of transmittingEscherichia coli O157: H7 and non-O157 to humans if they consume undercooked meat, being responsible for causing severe diseases such as hemolytic uremic syndrome. Bacterial resistance has become worrying concerning the efficacy in the treatment of diseases, and because of these aspects, the objective of this study was to verify the occurrence of E. coli O157: H7 and non-O157 in the stages of cattle slaughter and to evaluate the susceptibility of these bacteria against antimicrobial action. We collected from a cattle slaughterhouse 21 samples from manipulators' hands, 21 from knives and 300 from 50 animals in six points of the flowchart. The isolation was performed using the CT-SMAC agar and characterization of serotypes by PCR. There was higher occurrence of E. coli O157: H7 (12.0%) in animals and lower prevalence of E. coli O26 (8.0%) and O113 (2.0%). E. coli O26 was present in 9.52% of the knives. The presence of E. coli non-O157 sorbitol negative was an unexpected fact due to the method of isolation. All E. coli O157: H7 isolates were sensitive to tetracycline, cefepime, cefoxitin, ciprofloxacin and sulphazotrim, and 78.85% of them were resistant to cephalothin and 34.61% to ampicillin. All E. coli O26 were sensitive to cefepime, cefoxitin and sulphazotrim, and 88.23% were resistant to tetracycline and cephalothin and 82.35% to ampicillin. The antimicrobial multiresistance was observed in both serotypes. It should be, therefore, a criteria for using antimicrobial in treatments to avoid become a public health concern.

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Detection and Typing Strategies for Pathogenic Escherichia coli

Rivas¹

2015

SpringerBriefs in Food, Health, and Nutrition

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Pages: vi + 258 ISBN: Book: 978-1-910190-77-7. Ebook: 978-1-910190-78-4 £159, $319 Published by: Caister Academic Press www.caister.com Escherichia coli is an important member of the normal healthy microbiome of humans and other mammals. In addition, some strains are thought to be probiotic, and therefore beneficial to the host. However, other strains of E. coli have evolved into highly versatile, and frequently deadly, pathogens, the resultant diseases causing significant economic loss and public health burdens worldwide. Recent studies have shown that the E. coli genome has a high plasticity allowing it to adapt to new niches and survive in stressful conditions and to evolve into new hybrid strains with shared genes, including virulence genes. Omics and whole genome sequencing approaches have transformed research in this field allowing fascinating new insights into the molecular and cellular biology of the bacterium thus paving the way for the development of novel therapeutic strategies. Under the expert guidance of the editors in this book, renowned international authors provide timely and up-to-date reviews of current cutting-edge E. coli omics, molecular-and cellular-biology research. Topics range from E. coli genome plasticity and evolution to the application of omics technologies for in silico modeling to understand stress-triggered physiological responses. This authoritative volume is essential reading for scientists, both experts and students, working on pathogenic E. coli in academia, government, and biotechnology companies. It is also a must-read for anyone with an interest in bacterial pathogenesis and an important acquisition for all microbiology libraries.

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Applicability of a multiplex PCR to detect O26, O45, O103, O111, O121, O145, and O157 serogroups of Escherichia coli in cattle feces1

Cited by 73 publications

References 38 publications

Prevalence of Shiga Toxin–Producing Escherichia coli and Associated Virulence Genes in Feces of Commercial Feedlot Cattle

Prevalence of Shiga Toxin–Producing Escherichia coli and Associated Virulence Genes in Feces of Commercial Feedlot Cattle

Ocorrência de Escherichia coli O157:H7 e O26 sorbitol negativas em matadouro frigorífico de bovino e suscetibilidade a antimicrobianos

Detection and Typing Strategies for Pathogenic Escherichia coli

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