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Serotypes and virutypes of enteropathogenic and enterohaemorrhagic Escherichia coli strains from stool samples of children with diarrhoea in Germany
Authors:Kozub-Witkowski E  Krause G  Frankel G  Kramer D  Appel B  Beutin L
Affiliation: LADR GmbH, Medizinisches Versorgungszentrum, Dr. Kramer &Kollegen, Geesthacht, Germany;
 National Reference Laboratory for Escherichia coli;, Centre for Infectiology and Pathogen Characterization, Federal Institute for Risk Assessment, Berlin, Germany
 Division of Cell and Molecular Biology, Imperial College, London, UK;
 Department of Biological Safety, Federal Institute for Risk Assessment, Berlin, Germany
Abstract:Aims: To investigate the prevalence of traditional and emerging types of enteropathogenic (EPEC) and enterohaemorrhagic Escherichia coli (EHEC) strains in stool samples from children with diarrhoea and to characterize their virulence genes involved in the attaching and effacing (A/E) phenotype. Methods and Results: Serological and PCR‐based methods were used for detection and isolation of EPEC and EHEC strains from 861 stool samples from diarrhoeic children. Agglutination with traditional EPEC and EHEC O‐group‐specific antisera resulted in detection of 38 strains; 26 of these carried virulence factors of EPEC or EHEC. PCR screening for the eae gene resulted in isolation of 97 strains, five carried genes encoding Shiga toxins (stx), one carried the bfpA gene and 91 were atypical EPEC. The 97 EPEC and EHEC strains were divided into 36 O‐serogroups and 21 H‐types, only nine strains belonged to the traditional EPEC O‐groups O26, O55, O86 and O128. In contrast, EPEC serotypes O28:H28, O51:H49, O115:H38 and O127:H40 were found in multiple cases. Subtyping the virulence factors intimin, Tir and Tir‐cytoskeleton coupling effector protein (TccP)/TccP2 resulted in further classification of 93·8% of the 97 strains. Conclusions: Our findings show a clear advantage of the eae‐PCR over the serological detection method for identification of EPEC and EHEC strains from human patients. Significance and Impact of the Study: Molecular detection by the eae‐PCR followed by serotyping and virutyping is useful for monitoring trends in EPEC and EHEC infections and to discover their possible reservoirs.
Keywords:detection    diarrhoea    eae-PCR    enterohaemorrhagic E. coli    enteropathogenic E. coli    serotyping    virulence markers
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