Multiplex real-time PCR (MRT-PCR) for diarrheagenic

Francesca Barletta; Theresa J. Ochoa; Thomas G. Cleary

doi:10.1007/978-1-60327-353-4_21

Multiplex real-time PCR (MRT-PCR) for diarrheagenic

Francesca Barletta
, Theresa J. Ochoa
, Thomas G. Cleary

School of Medicine

Universidad Peruana Cayetano Heredia, Instituto de Medicina Tropical Alexander von Humboldt
University of Texas Health Science Center at Houston

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

20 Scopus citations

Abstract

Diarrheagenic Escherichia coli strains are important causes of diarrhea in children from the developing world and are now being recognized as emerging enteropathogens in the developed world. Current methods of detection are too expensive and labor-intensive for routine detection of these organisms to be practical. We developed a real-time fluorescence-based multiplex PCR for the detection of all six of the currently recognized classes of diarrheagenic E. coli. The primers were designed to specifically amplify eight different virulence genes in the same reaction: aggR for enteroaggregative E. coli (EAEC), stIa/stIb and lt for enterotoxigenic E. coli (ETEC), eaeA for enteropathogenic E. coli (EPEC), stx1 and stx2 for Shiga toxin-producing E. coli (STEC), ipaH for enteroinvasive E. coli (EIEC), and daaD for diffusely adherent E. coli (DAEC).

Original language	English
Title of host publication	PCR Detection of Microbial Pathogens
Editors	Mark Wilks
Pages	307-314
Number of pages	8
DOIs	https://doi.org/10.1007/978-1-60327-353-4_21
State	Published - 2013

Publication series

Name	Methods in Molecular Biology
Volume	943
ISSN (Print)	1064-3745

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

Diarrhea
Diarrheagenic E. coli
Diffusely adherent E. coli
Enteroaggregative E. coli
Enteroinvasive E. coli
Enteropathogenic E. coli
Enterotoxigenic E. coli
Multiplex real-time PCR
Shiga toxin-producing E. coli

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10.1007/978-1-60327-353-4_21

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abstract = "Diarrheagenic Escherichia coli strains are important causes of diarrhea in children from the developing world and are now being recognized as emerging enteropathogens in the developed world. Current methods of detection are too expensive and labor-intensive for routine detection of these organisms to be practical. We developed a real-time fluorescence-based multiplex PCR for the detection of all six of the currently recognized classes of diarrheagenic E. coli. The primers were designed to specifically amplify eight different virulence genes in the same reaction: aggR for enteroaggregative E. coli (EAEC), stIa/stIb and lt for enterotoxigenic E. coli (ETEC), eaeA for enteropathogenic E. coli (EPEC), stx1 and stx2 for Shiga toxin-producing E. coli (STEC), ipaH for enteroinvasive E. coli (EIEC), and daaD for diffusely adherent E. coli (DAEC).",

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AU - Cleary, Thomas G.

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N2 - Diarrheagenic Escherichia coli strains are important causes of diarrhea in children from the developing world and are now being recognized as emerging enteropathogens in the developed world. Current methods of detection are too expensive and labor-intensive for routine detection of these organisms to be practical. We developed a real-time fluorescence-based multiplex PCR for the detection of all six of the currently recognized classes of diarrheagenic E. coli. The primers were designed to specifically amplify eight different virulence genes in the same reaction: aggR for enteroaggregative E. coli (EAEC), stIa/stIb and lt for enterotoxigenic E. coli (ETEC), eaeA for enteropathogenic E. coli (EPEC), stx1 and stx2 for Shiga toxin-producing E. coli (STEC), ipaH for enteroinvasive E. coli (EIEC), and daaD for diffusely adherent E. coli (DAEC).

AB - Diarrheagenic Escherichia coli strains are important causes of diarrhea in children from the developing world and are now being recognized as emerging enteropathogens in the developed world. Current methods of detection are too expensive and labor-intensive for routine detection of these organisms to be practical. We developed a real-time fluorescence-based multiplex PCR for the detection of all six of the currently recognized classes of diarrheagenic E. coli. The primers were designed to specifically amplify eight different virulence genes in the same reaction: aggR for enteroaggregative E. coli (EAEC), stIa/stIb and lt for enterotoxigenic E. coli (ETEC), eaeA for enteropathogenic E. coli (EPEC), stx1 and stx2 for Shiga toxin-producing E. coli (STEC), ipaH for enteroinvasive E. coli (EIEC), and daaD for diffusely adherent E. coli (DAEC).

KW - Diarrhea

KW - Diarrheagenic E. coli

KW - Diffusely adherent E. coli

KW - Enteroaggregative E. coli

KW - Enteroinvasive E. coli

KW - Enteropathogenic E. coli

KW - Enterotoxigenic E. coli

KW - Multiplex real-time PCR

KW - Shiga toxin-producing E. coli

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U2 - 10.1007/978-1-60327-353-4_21

DO - 10.1007/978-1-60327-353-4_21

M3 - Capítulo

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AN - SCOPUS:84870549857

SN - 9781603273527

T3 - Methods in Molecular Biology

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EP - 314

BT - PCR Detection of Microbial Pathogens

A2 - Wilks, Mark

ER -

Multiplex real-time PCR (MRT-PCR) for diarrheagenic

Abstract

Publication series

UN SDGs

Keywords

Access to Document

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