Rapid detection of diarrheagenic Escherichia coli by Real Time PCR

• Grant number: 08/11467-9
• Support Opportunities: Regular Research Grants
• Start date: April 01, 2009
• End date: September 30, 2011
• Field of knowledge: Biological Sciences - Microbiology - Biology and Physiology of Microorganisms
• Principal Investigator: Isabel Cristina Affonso Scaletsky
• Grantee: Isabel Cristina Affonso Scaletsky
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil

Abstract

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)