ArticleJournal of clinical laboratory analysis2013
Detection of diarrheagenic Escherichia coli using a two-system multiplex-PCR protocol.
Article in Journal of clinical laboratory analysis, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed, 19 citations in OpenAlex.
- Intestinal and Extraintestinal Pathotypes ofAntibiotics (Basel, Switzerland) · 2025Article
- [Distribution of diarrheagenicRevista espanola de quimioterapia : publicacion oficial de la Sociedad Espanola de Quimioterapia · 2025Article
- Genomic epidemiology of extended-spectrum beta-lactamase-producingMicrobial genomics · 2025Article
- Article
- PCR diagnostics are insufficient for the detection of Diarrhoeagenic Escherichia coli in Ibadan, Nigeria.PLOS global public health · 2023Article
- Article
- Increased sensitivity of enterotoxigenicBiotechnology reports (Amsterdam, Netherlands) · 2021Article
- Distinguishing Pathovars from Nonpathovars: Escherichia coli.Microbiology spectrum · 2020Review
- Detection and discrimination of five E. coli pathotypes using a combinatory SYBR® Green qPCR screening system.Applied microbiology and biotechnology · 2018Article
- Development and accuracy of quantitative real-time polymerase chain reaction assays for detection and quantification of enterotoxigenic Escherichia coli (ETEC) heat labile and heat stable toxin genes in travelers' diarrhea samples.The American journal of tropical medicine and hygiene · 2014Article
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Authors and funding
11 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundDiarrheagenic Escherichia coli (DEC) strains are important causes of diarrhea. However, they cannot be distinguished from E. coli of the intestinal microbiota by conventional microbiological tests.
methodsThis work presents a two-system multiplex PCR for detection of DEC. Primers for 16S rRNA gene were added as internal amplification control to validate negative reactions. The multiplex-PCR system 1 contains primers for detection of Shiga toxin producing E. coli (STEC; stx1, stx2), enteropathogenic E. coli (EPEC; eae, bfpA), atypical enteropathogenic E. coli (aEPEc; eae), enteroinvasive E. coli (ETEC; lt, st), enteroinvasive E. coli (EIEC; ial), and the internal amplification control 16S rRNA. The system 2 contains primers for EIEC (ipaH), enteroaggregative E. coli (CVD432), diffusely adherent E. coli (daaE), and 16S rRNA. The protocol was tested with E. coli reference strains, and also with cultures of fecal specimens of people with diarrhea and healthy controls.
resultsThe protocol correctly identified the DEC reference strains. No DEC marker was amplified for negative controls; these results were validated by the amplification of a fragment of the 16S rRNA gene. The frequency of DEC was 7.6% for both patients and healthy controls; two Shigella sonnei strains were detected in the group with diarrhea. The identity of the amplicons was confirmed by DNA sequencing.
conclusionThe protocol is specific for DEC Shigella and is suitable for clinical laboratories.
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