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Evaluation of the 'GeneDisc' real-time PCR system for detection of enterohaemorrhagic Escherichia coli (EHEC) O26, O103, O111, O145 and O157 strains according to their virulence markers and their O- and H-antigen-associated genes.
MedLine Citation:
PMID:  19191965     Owner:  NLM     Status:  In-Process    
Abstract/OtherAbstract:
AIMS: To evaluate the GeneDisc multiplex real-time PCR assay for detection of enterohaemorrhagic Escherichia coli (EHEC) O26, O103, O111, O145 and O157 strains. METHODS AND RESULTS: GeneDiscs for detection of genes encoding Shiga toxins (stx), intimins (eae), E. coli O157 (rfbE(O157)) and H7 (fliC(H7)) antigens as well as genes specific for EHEC O26 (wzx(O26)), O103 (wzx(O103)), O111 (wbd1(O111)), O145 (ihp1(O145)) and O157 (ihp1(O157)) were evaluated. The assay was run with native bacteria in 1 h in a GeneDisc Cycler. All genotypes of stx and eae, except stx(2f) and eae-rho, were identified. Escherichia coli strains belonging to O-groups O26, O103, O111, O157 as well as EHEC O145:[H28] strains were specifically detected with this assay. The ihp1(O157) gene was not found specific for EHEC O157. O-rough mutants of EHEC and non-motile EHEC O157 strains were reliably identified with the GeneDisc assay. Two to three colonies of EHEC strains were still detectable in a lawn of 50 000 apathogenic E. coli from agar plates. CONCLUSIONS: The GeneDisc assay is a specific and reliable assay for detection of major EHEC strains. It is robust enough to detect few EHEC colonies in mixed cultures of bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: The assay is promising for its use in EHEC diagnostics and for EHEC monitoring with different kinds of samples.
Authors:
L Beutin; S Jahn; P Fach
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Publication Detail:
Type:  Journal Article     Date:  2009-01-15
Journal Detail:
Title:  Journal of applied microbiology     Volume:  106     ISSN:  1365-2672     ISO Abbreviation:  J. Appl. Microbiol.     Publication Date:  2009 Apr 
Date Detail:
Created Date:  2009-03-17     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9706280     Medline TA:  J Appl Microbiol     Country:  England    
Other Details:
Languages:  eng     Pagination:  1122-32     Citation Subset:  IM    
Affiliation:
Centre for Infectiology and Pathogen Characterization, Federal Institute for Risk Assessment, Berlin, Germany. lothar.beutin@bfr.bund.de
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