A method for fast and simple detection of major diarrhoeagenic Escherichia coli in the routine diagnostic laboratory
作者： S. Persson , K.E.P. Olsen , F. Scheutz , K.A. Krogfelt , P. Gerner-Smidt
DOI: 10.1111/J.1469-0691.2007.01692.X
关键词: Shigella 、 Virulence 、 Multiplex polymerase chain reaction 、 Enterotoxin 、 Biology 、 Verocytotoxin 、 Microbiology 、 Escherichia coli 、 VTEC 、 Molecular biology 、 Intimin
摘要: A multiplex PCR was developed for the detection of following genes characteristic diarrhoeagenic Escherichia coli (DEC): verocytotoxins 1 (vtx1) and 2 (vtx2), verocytotoxin-producing E. (VTEC); intimin (eae), found in enteropathogenic (EPEC), attaching effacing VTEC; heat-stable enterotoxin (estA) heat-labile (eltA), enterotoxigenic (ETEC); invasive plasmid antigen (ipaH), enteroinvasive (EIEC) Shigella spp. The method allowed simultaneous identification all six one reaction, included a 16S rDNA internal control. When applied to pure cultures from reference strain collection, virulence 124 different DEC strains 15 were identified correctly, there no cross-reactions with 13 non-E. species. limit 10(2)-10(3) CFU/PCR presence 10(6) non-target cells. tested colonies plate clinical stool samples, it faster, more sensitive, less expensive laborious diagnostic procedure than DNA hybridisation. used purified spiked samples (by two commercial kits), had CFU/mL sample.
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