Detection of Escherichia coli, Salmonella spp., Shigella spp., Yersinia enterocolitica, Vibrio cholerae, and Campylobacter spp. enteropathogens by 3-reaction multiplex polymerase chain reaction

Oscar G Gómez-Duarte; Jing Bai; Elizabeth Newell

doi:10.1016/j.diagmicrobio.2008.09.006

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Detection of Escherichia coli, Salmonella spp., Shigella spp., Yersinia enterocolitica, Vibrio cholerae, and Campylobacter spp. enteropathogens by 3-reaction multiplex polymerase chain reaction

Journal article

Peer reviewed

Detection of Escherichia coli, Salmonella spp., Shigella spp., Yersinia enterocolitica, Vibrio cholerae, and Campylobacter spp. enteropathogens by 3-reaction multiplex polymerase chain reaction

Oscar G Gómez-Duarte, Jing Bai and Elizabeth Newell

Diagnostic microbiology and infectious disease, Vol.63(1), pp.1-9

01/2009

DOI: 10.1016/j.diagmicrobio.2008.09.006

PMCID: PMC2701628

PMID: 18990527

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Abstract

The magnitude of bacterial diarrhea in developing countries is largely unknown because affordable detection methods are not available. We have developed a polymerase chain reaction (PCR)-based assay for use in areas with limited resources to screen for diarrheogenic strains from clinical isolates. To simplify the assay and minimize reagents, our method implemented the use of plasmids rather than bacteria as template controls and the use of bacterial suspensions or crude DNA preparations rather than purified genomic DNA as template DNA. The assay consisted of 3 PCR reactions using 3 groups of 5 to 6 primer pairs to identify the 11 most common bacterial diarrheogenic pathogens. The 3-reaction multiplex PCR amplifies DNA targets specific for each 1 of the 6 Escherichia coli diarrheogenic strains and the 5 non-E. coli diarrheogenic strains, including Salmonella spp., Shigella spp., Campylobacter spp., Yersinia enterocolitica, and Vibrio cholerae. The assay may provide an important epidemiologic tool to investigate the role of diarrheogenic bacterial pathogens in areas of the world with limited resources.

Reproducibility of Results

Genes, Bacterial - genetics

Vibrio cholerae - genetics

Humans

Enterobacteriaceae - genetics

Salmonella - genetics

DNA Primers - genetics

Enterobacteriaceae - isolation & purification

Shigella - isolation & purification

Yersinia enterocolitica - isolation & purification

Campylobacter - genetics

Enteropathogenic Escherichia coli - genetics

Polymerase Chain Reaction - methods

Diarrhea - microbiology

Shigella - genetics

Electrophoresis, Agar Gel

Salmonella - isolation & purification

Yersinia enterocolitica - genetics

Sensitivity and Specificity

Cloning, Molecular

Campylobacter - isolation & purification

Developing Countries

Enteropathogenic Escherichia coli - isolation & purification

Vibrio cholerae - isolation & purification

Details

Title: Subtitle: Detection of Escherichia coli, Salmonella spp., Shigella spp., Yersinia enterocolitica, Vibrio cholerae, and Campylobacter spp. enteropathogens by 3-reaction multiplex polymerase chain reaction
Creators: Oscar G Gómez-Duarte - Division of Pediatric Infectious Diseases, Department of Pediatrics, University of Iowa Children's Hospital, Iowa City, IA 52242, USA. oscar-gomez@uiowa.edu
Jing Bai
Elizabeth Newell
Resource Type: Journal article
Publication Details: Diagnostic microbiology and infectious disease, Vol.63(1), pp.1-9
DOI: 10.1016/j.diagmicrobio.2008.09.006
PMID: 18990527
PMCID: PMC2701628
NLM abbreviation: Diagn Microbiol Infect Dis
ISSN: 0732-8893
eISSN: 1879-0070
Publisher: United States
Grant note: K12 HD027748-16A1 / NICHD NIH HHS
Language: English
Date published: 01/2009
Academic Unit: Critical Care; Stead Family Department of Pediatrics; Iowa Neuroscience Institute
Record Identifier: 9984070410702771

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