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dc.contributor.author
Botkin, Douglas J.
dc.contributor.author
Galli, Lucía
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dc.contributor.author
Sankarapani, Vinoth
dc.contributor.author
Soler, Michael
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Rivas, Marta
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dc.contributor.author
Torres, Alfredo G.
dc.date.available
2019-05-30T23:13:35Z
dc.date.issued
2012-02
dc.identifier.citation
Botkin, Douglas J.; Galli, Lucía; Sankarapani, Vinoth; Soler, Michael; Rivas, Marta; et al.; Development of a Multiplex PCR Assay for Detection of Shiga Toxin-Producing Escherichia coli, Enterohemorrhagic E. coli, and Enteropathogenic E. coli Strains; Frontiers Research Foundation; Frontiers in Cellular and Infection Microbiology; 2; 8; 2-2012; 1-10
dc.identifier.issn
1664-302X
dc.identifier.uri
http://hdl.handle.net/11336/77434
dc.description.abstract
Escherichia coli O157:H7 and other pathogenic E.coli strains are enteric pathogens associated with food safety threats and which remaina significant cause of morbidity and mortality worldwide. In the current study, we investigated whether enterohemorrhagic E. coli (EHEC), Shiga toxin-producing E. coli (STEC), and enteropathogenic E. coli (EPEC) strains can be rapidly and specifically differentiated with multiplex PCR (mPCR) utilizing selected biomarkers associated with each strains respective virulence genotype. Primers were designed to amplify multiple intimin (eae) and long polar fimbriae (lpfA) variants, the bundle forming pilus gene bfpA, and the Shiga toxin encoding genes stx1 and stx2. We demonstrated consistent amplification of genes specific to the prototype EHEC O157:H7 EDL933 (lpfA1-3, lpfA2-2, stx1, stx2, and eae-gama) and EPEC O127:H6 E2348/69 (eae-alfa, lpfA1-1, and bfpA) strains using the optimized mPCR protocol with purified genomic DNA (gDNA). A screen of gDNA from isolates in a diarrheagenic E. coli collection revealed that the mPCR assay was successful in predicting the correct pathotype of EPEC and EHEC clones grouped in the distinctive phyogenetic disease clusters EPEC1 and EHEC1, and was able to differentiate EHEC1 from EHEC2 clusters. The assay detection threshold was 2×104 CFU per PCR reaction for EHEC and EPEC. mPCR was also used to screen Argentinean clinical samples from hemolytic uremic syndrome and diarrheal patients, resultingin 91 % sensitivity and 84% specificity when compared to established molecular diagnostic procedures. In conclusion, our mPCR methodology permitted differentiation of EPEC, STEC and EHEC strains from other pathogenic E.coli; therefore, the assay becomes an additional tool for rapid diagnosis of these organisms.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Frontiers Research Foundation
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dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc/2.5/ar/
dc.subject
Stec
dc.subject
Etec
dc.subject
Epec
dc.subject
Diagnostics
dc.subject.classification
Salud Ocupacional
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dc.subject.classification
Ciencias de la Salud
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dc.subject.classification
CIENCIAS MÉDICAS Y DE LA SALUD
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dc.title
Development of a Multiplex PCR Assay for Detection of Shiga Toxin-Producing Escherichia coli, Enterohemorrhagic E. coli, and Enteropathogenic E. coli Strains
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2019-05-08T17:21:50Z
dc.identifier.eissn
2235-2988
dc.journal.volume
2
dc.journal.number
8
dc.journal.pagination
1-10
dc.journal.pais
Suiza
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dc.journal.ciudad
Lausana
dc.description.fil
Fil: Botkin, Douglas J.. University of Texas Medical Branch; Estados Unidos
dc.description.fil
Fil: Galli, Lucía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”; Argentina. University of Texas Medical Branch; Estados Unidos
dc.description.fil
Fil: Sankarapani, Vinoth. University of Texas Medical Branch; Estados Unidos
dc.description.fil
Fil: Soler, Michael. University of Texas Medical Branch; Estados Unidos
dc.description.fil
Fil: Rivas, Marta. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”; Argentina
dc.description.fil
Fil: Torres, Alfredo G.. University of Texas Medical Branch; Estados Unidos
dc.journal.title
Frontiers in Cellular and Infection Microbiology
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3417533/
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://www.frontiersin.org/articles/10.3389/fcimb.2012.00008/full
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/https://doi.org/10.3389/fcimb.2012.00008
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