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Rapid detection of diarrheagenic Escherichia coli by Real Time PCR

Grant number: 08/11467-9
Support type:Regular Research Grants
Duration: April 01, 2009 - September 30, 2011
Field of knowledge:Biological Sciences - Microbiology
Principal Investigator:Isabel Cristina Affonso Scaletsky
Grantee:Isabel Cristina Affonso Scaletsky
Home Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil


Diarrheagenic Escherichia coli (DEC) strains were among the most important causes of childhood diarrhea. These DEC strains can be classified into five major categories on the basis of the nature of their infection and pathogenic mechanisms: enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), enterohemorrhagic E. coli (EHEC) and enteroaggregative E. coli (EAEC). DEC strains were among the first pathogens for which molecular diagnostic methods were developed. PCR-based methods are more sensitive and rapid than phenotypes tests performed on individual colonies. Recently, we have developed multiplex PCR assays to detect EPEC, EAEC, ETEC, EIEC, and EHEC isolates in stool specimens after culture on MacConkey agar. The aim of this project is to adapt these protocols to real-time PCR for the rapid detection of DEC, eliminating the need for gel electrophoresis and risk for carry-over contamination. (AU)