Detection of Diarrheagenic Escherichia coli Using a Two-System Multiplex-PCR Protocol. Fialho, O., B., de Souza, E., M., de Borba Dallagassa, C., de Oliveira Pedrosa, F., Klassen, G., Irino, K., Paludo, K., S., de Assis, F., E., A., Surek, M., de Souza Santos Farah, S., M., & Fadel-Picheth, C., M., T. Journal of Clinical Laboratory Analysis, 27(2):155-161, Wiley-Blackwell, 3, 2013. ![Detection of Diarrheagenic Escherichia coli Using a Two-System Multiplex-PCR Protocol [link]](https://bibbase.org/img/filetypes/link.svg "link")
Website abstract bibtex BACKGROUND: Diarrheagenic 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. METHODS: This 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. RESULTS: The 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. CONCLUSION: The protocol is specific for DEC Shigella and is suitable for clinical laboratories.
@article{
title = {Detection of Diarrheagenic Escherichia coli Using a Two-System Multiplex-PCR Protocol},
type = {article},
year = {2013},
identifiers = {[object Object]},
keywords = {Diarrhea,Enteropathogens,Internal PCR control,Molecular diagnosis,Molecular method},
pages = {155-161},
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websites = {http://doi.wiley.com/10.1002/jcla.21578},
month = {3},
publisher = {Wiley-Blackwell},
id = {0e7a4e83-a127-3d90-b69f-b6a9475627bc},
created = {2019-02-04T15:29:25.258Z},
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abstract = {BACKGROUND: Diarrheagenic 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. METHODS: This 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. RESULTS: The 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. CONCLUSION: The protocol is specific for DEC Shigella and is suitable for clinical laboratories.},
bibtype = {article},
author = {Fialho, Octaviana Baccin and de Souza, Emanuel Maltempi and de Borba Dallagassa, Cibelle and de Oliveira Pedrosa, Fábio and Klassen, Giseli and Irino, Kinue and Paludo, Katia Sabrina and de Assis, Flávia Emanoelli Araújo and Surek, Monica and de Souza Santos Farah, Sônia Maria and Fadel-Picheth, Cyntia Maria Telles},
journal = {Journal of Clinical Laboratory Analysis},
number = {2}
}
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