Evaluation of a multiplex PCR method to detect enteroaggregative Escherichia coli

Enteroaggregative Escherichia coli (EAEC) has been implicated in sporadic diarrhea in children and adults and has been identified as the cause of several outbreaks worldwide. The HEp-2 test remains the gold standard for identification of this pathotype. A 60-65 MDa plasmid encodes the aggregative ad...

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Bibliographic Details
Published in:	Biocell Vol. 30; no. 2; pp. 301 - 308
Main Authors:	Rüttler, M E, Yanzón, C S, Cuitiño, M J, Renna, N F, Pizarro, M A, Ortiz, A M
Format:	Journal Article
Language:	English
Published:	Argentina Tech Science Press 01-01-2006 Sociedad Latinoamericana de Microscopía Electrónica.; Centro Regional de Investigaciones Científicas y Tecnológicas (Mendoza, Argentina)
Subjects:	Bacterial Adhesion - physiology BIOLOGY Children Diarrhea DNA, Bacterial - analysis DNA, Bacterial - genetics E coli Electrophoresis, Agar Gel Escherichia coli Escherichia coli - cytology Escherichia coli - genetics Escherichia coli - isolation & purification Escherichia coli Proteins - genetics Humans Infant Pili Polymerase chain reaction Polymerase Chain Reaction - methods Serotyping Thermal stability Trans-Activators - genetics Transcription Diarrhea Cell culture PCR (Polymerase Chain Reaction) Escherichia coli
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Summary:	Enteroaggregative Escherichia coli (EAEC) has been implicated in sporadic diarrhea in children and adults and has been identified as the cause of several outbreaks worldwide. The HEp-2 test remains the gold standard for identification of this pathotype. A 60-65 MDa plasmid encodes the aggregative adherence fimbriae (AAF/I and AAF/II), a transcriptional activator (aggR gene), the enteroaggregative heat-stable enterotoxin EAST1 (astA gene) and a cytotoxin (Pet). The standard assay for EAEC is performed only in research laboratories, because it is expensive, labor intensive and time-consuming. The Polymerase Chain Reaction (PCR) offers the possibility of rapid diagnosis. In the current study, a multiplex PCR assay which checks aggR and astA genes was designed. Eigthy-eight E. coli strains, isolated from children with acute diarrhea in Mendoza, Argentina, were characterized by the reference method (HEp-2 assay), and by aggR-astA PCR. A strong correlation between the presence of the specific marker aggR and the reference test was found. The astA gene had a similar distribution between aggregative and localized strains, indicating that this gene could not be considered as a marker of EAEC. We conclude that aggR may be used to identify EAEC, using the PCR method as a screening test.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0327-9545 1667-5746
DOI:	10.32604/biocell.2006.30.301