Detection of Escherichia coli O157:H7 by multiplex PCR.
In order to develop a PCR assay for Escherichia coli O157:H7, a portion of the 60-MDa plasmid harbored by enterohemorrhagic E. coli (EHEC) was sequenced and PCR primers were designed. A multiplex PCR method was then designed by employing primers specific for the EHEC eaeA gene, conserved sequences of Shiga-like toxins I (SLT-I) and II (SLT-II), and the 60-MDa plasmid. PCR products of 1,087 bp (eaeA), 227 and/or 224 bp (SLT-I and/or SLT-II), and 166 bp (plasmid) were successfully amplified simultaneously in a single reaction. The multiplex PCR method can be used to specifically identify EHEC of serogroup O157.
Fratamico
P M
PM
Eastern Regional Research Center, U.S. Department of Agriculture, Philadelphia, Pennsylvania 19118, USA.
Sackitey
S K
SK
Wiedmann
M
M
Deng
M Y
MY
D001427:Bacterial Toxins / Q000235:genetics; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D015342:DNA Probes / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000145:classification* / Q000235:genetics* / Q000472:pathogenicity; D004927:Escherichia coli Infections / Q000382:microbiology; D005069:Evaluation Studies as Topic; D005798:Genes, Bacterial; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012703:Serotyping; D022622:Shiga Toxin 1; D022641:Shiga Toxin 2; D013045:Species Specificity; D014774:Virulence / Q000235:genetics
D001427:Bacterial Toxins; D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial; D022622:Shiga Toxin 1; D022641:Shiga Toxin 2