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Journal of Clinical Microbiology, February 1998, p. 598-602, Vol. 36, No. 2
Molecular Microbiology Unit, Women's and
Children's Hospital, North Adelaide, South Australia 5006, Australia
Received 16 June 1997/Returned for modification 21 October
1997/Accepted 18 November 1997
Shiga toxigenic Escherichia coli (STEC) comprises a
diverse group of organisms capable of causing severe gastrointestinal disease in humans. Within the STEC family, certain strains appear to be
of greater virulence for humans, for example, those belonging to
serogroups O111 and O157 and those with particular combinations of
other putative virulence traits. We have developed two multiplex PCR
assays for the detection and genetic characterization of STEC in
cultures of feces or foodstuffs. Assay 1 utilizes four PCR primer pairs
and detects the presence of stx1,
stx2 (including variants of
stx2), eaeA, and enterohemorrhagic
E. coli hlyA, generating amplification products of 180, 255, 384, and 534 bp, respectively. Assay 2 uses two primer pairs
specific for portions of the rfb (O-antigen-encoding)
regions of E. coli serotypes O157 and O111, generating PCR
products of 259 and 406 bp, respectively. The two assays were validated
by testing 52 previously characterized STEC strains and observing 100%
agreement with previous results. Moreover, assay 2 did not give a
false-positive O157 reaction with enteropathogenic E. coli
strains belonging to clonally related serogroup O55. Assays 1 and 2 detected STEC of the appropriate genotype in primary fecal cultures
from five patients with hemolytic-uremic syndrome and three with bloody
diarrhea. Thirty-one other primary fecal cultures from patients without
evidence of STEC infection were negative.
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Detection and Characterization of Shiga Toxigenic
Escherichia coli by Using Multiplex PCR Assays for
stx1, stx2,
eaeA, Enterohemorrhagic E. coli hlyA,
rfbO111, and
rfbO157
*
Corresponding author. Mailing address: Molecular
Microbiology Unit, Women's and Children's Hospital, North Adelaide,
S.A. 5006, Australia. Phone: 61-8-8204 6302. Fax: 61-8-8204 6051. E-mail: patonj{at}wch.sa.gov.au.
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