title: Detection by 5'-nuclease PCR of Shiga-toxin producing Escherichia coli O26, O55, O91, O103, O111, O113, O145 and O157:H7, associated with the world's most frequent clinical cases.
abstract: This paper describes 5'-nuclease PCR assays for detecting eight O-serogroups, H7 flagellar antigen and stx genes from the Shiga toxin-producing Escherichia coli (STEC) associated with the world's most frequent clinical cases. A single set of primers was used to detect the genes stx1 and stx2 in the same reaction by 5'-nuclease PCR. Serotyping by 5'-nuclease PCR of STEC was based on the selection of primers and probes targeting the O-antigen gene clusters of E. coli O26, O55, O91, O111, O113, O157, the eae gene of E. coli O103, the O-island 29 of E. coli O145, and the flagellar H7 antigen gene. Results obtained on a collection of 190 strains indicate that the 5'-nuclease PCR assays used here could serve as a basis for rapid specific stx, O and H7 typing of these major pathogenic serogroups of E. coli. This work provides sensitive and specific tests for the rapid, reliable detection of the main pathogenic E. coli O-serogroups of major public health concern.
authors.affiliation: Agence Française de Sécurité Sanitaire des Aliments (AFSSA), Laboratoire d'Etudes et de Recherches sur la Qualité des Aliments et sur les Procédés Agroalimentaires, Unité: Etude moléculaire des contaminants biologiques alimentaires, Maisons-Alfort, France.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Perelle
authors.forename: Françoise
authors.initials: F
authors.lastname: Dilasser
authors.forename: Joël
authors.initials: J
authors.lastname: Grout
authors.forename: Patrick
authors.initials: P
authors.lastname: Fach
mesh_terms: D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000145:classification / Q000235:genetics* / Q000472:pathogenicity*; D004927:Escherichia coli Infections / Q000382:microbiology*; D019453:Escherichia coli O157 / Q000145:classification / Q000235:genetics / Q000472:pathogenicity; D005798:Genes, Bacterial / Q000235:genetics; D043264:Phosphodiesterase I / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012703:Serotyping; D022622:Shiga Toxin 1 / Q000235:genetics; D022641:Shiga Toxin 2 / Q000235:genetics; D022603:Shiga Toxins / Q000235:genetics*
chemical_list: D017931:DNA Primers; D022622:Shiga Toxin 1; D022641:Shiga Toxin 2; D022603:Shiga Toxins; D043264:Phosphodiesterase I