Biomaterial Database

Multiplex polymerase chain reaction typing scheme based on Escherichia coli O157:H7 Sakai prophage (Sp)-associated genes. 2022

István Tóth, and Eva Bagyinszky, and Domonkos Sváb

Veterinary Medical Research Institute, Hungária krt. 21., H-1143, Budapest, Hungary.

OBJECTIVE Escherichia coli strains of the O157 serogroup include significant foodborne pathogens: enterohemorrhagic E. coli (EHEC) and enteropathogenic E. coli, which are responsible for a considerable number of hospitalizations and deaths worldwide each year. There is a constant need for rapid, reliable, and easy-to-use methods for their identification, typing, and phylogenetic classification. In this study, we proposed a new multiplex polymerase chain reaction (PCR)-based typing system for pathogenic E. coli, focusing on the O157 serogroup. METHODS We designed primers targeting 12 lambdoid prophage regions carried by the prototypic polylysogenic strain of EHEC, the O157:H7 Sakai strain. The reactions were tested in vitro as well as in silico with the PubMLST database. RESULTS The PCR assays can be grouped into four multiplex reactions, and their results can be given as a four-digit code. In vitro and in silico testing showed that these Sakai prophage regions are prevalent not only in E. coli O157 strains but also in Shiga toxigenic E. coli non-O157 strains and the method provides appropriate resolution. CONCLUSIONS The proposed method could be a valuable tool in epidemiologic tracing and preliminary phylogenetic grouping of this diverse group of pathogens.

UI	MeSH Term	Description	Entries
D010802	Phylogeny	The relationships of groups of organisms as reflected by their genetic makeup.	Community Phylogenetics,Molecular Phylogenetics,Phylogenetic Analyses,Phylogenetic Analysis,Phylogenetic Clustering,Phylogenetic Comparative Analysis,Phylogenetic Comparative Methods,Phylogenetic Distance,Phylogenetic Generalized Least Squares,Phylogenetic Groups,Phylogenetic Incongruence,Phylogenetic Inference,Phylogenetic Networks,Phylogenetic Reconstruction,Phylogenetic Relatedness,Phylogenetic Relationships,Phylogenetic Signal,Phylogenetic Structure,Phylogenetic Tree,Phylogenetic Trees,Phylogenomics,Analyse, Phylogenetic,Analysis, Phylogenetic,Analysis, Phylogenetic Comparative,Clustering, Phylogenetic,Community Phylogenetic,Comparative Analysis, Phylogenetic,Comparative Method, Phylogenetic,Distance, Phylogenetic,Group, Phylogenetic,Incongruence, Phylogenetic,Inference, Phylogenetic,Method, Phylogenetic Comparative,Molecular Phylogenetic,Network, Phylogenetic,Phylogenetic Analyse,Phylogenetic Clusterings,Phylogenetic Comparative Analyses,Phylogenetic Comparative Method,Phylogenetic Distances,Phylogenetic Group,Phylogenetic Incongruences,Phylogenetic Inferences,Phylogenetic Network,Phylogenetic Reconstructions,Phylogenetic Relatednesses,Phylogenetic Relationship,Phylogenetic Signals,Phylogenetic Structures,Phylogenetic, Community,Phylogenetic, Molecular,Phylogenies,Phylogenomic,Reconstruction, Phylogenetic,Relatedness, Phylogenetic,Relationship, Phylogenetic,Signal, Phylogenetic,Structure, Phylogenetic,Tree, Phylogenetic
D004927	Escherichia coli Infections	Infections with bacteria of the species ESCHERICHIA COLI.	E coli Infections,E. coli Infection,Infections, E coli,Infections, Escherichia coli,E coli Infection,E. coli Infections,Escherichia coli Infection,Infection, E coli,Infection, E. coli,Infection, Escherichia coli
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D054323	Shiga-Toxigenic Escherichia coli	Strains of ESCHERICHIA COLI with the ability to produce at least one or more of at least two antigenically distinct, usually bacteriophage-mediated cytotoxins: SHIGA TOXIN 1 and SHIGA TOXIN 2. These bacteria can cause severe disease in humans including bloody DIARRHEA and HEMOLYTIC UREMIC SYNDROME.	E coli, Verotoxigenic,Escherichia coli, Verotoxigenic,STEC,Verotoxigenic Escherichia coli,Shiga Toxigenic E. coli,Shiga Toxigenic Escherichia coli,Shiga Toxin-Producing Escherichia coli,VTEC,Vero Cytotoxin-Producing Escherichia coli,Verotoxigenic E. coli,Verotoxin-Producing Escherichia coli
D054324	Enterohemorrhagic Escherichia coli	Strains of ESCHERICHIA COLI that are a subgroup of SHIGA-TOXIGENIC ESCHERICHIA COLI. They cause non-bloody and bloody DIARRHEA; HEMOLYTIC UREMIC SYNDROME; and hemorrhagic COLITIS. An important member of this subgroup is ESCHERICHIA COLI O157-H7.	E coli, Enterohemorrhagic,EHEC,Escherichia coli, Enterohemorrhagic,Enterohemorrhagic E coli,Enterohemorrhagic E. coli
D060885	Multiplex Polymerase Chain Reaction	Methods for using more than one primer set in a polymerase chain reaction to amplify more than one segment of the target DNA sequence in a single reaction.	Multiplex Ligation-Dependent Probe Amplification,Multiplex PCR,Triplex PCR,Triplex Polymerase Chain Reaction,Multiplex Ligation Dependent Probe Amplification,PCR, Multiplex,PCR, Triplex
D019453	Escherichia coli O157	A verocytotoxin-producing serogroup belonging to the O subfamily of Escherichia coli which has been shown to cause severe food-borne disease. A strain from this serogroup, serotype H7, which produces SHIGA TOXINS, has been linked to human disease outbreaks resulting from contamination of foods by E. coli O157 from bovine origin.	E coli O157,E coli O157-H7,Escherichia coli O157-H7
D029968	Escherichia coli Proteins	Proteins obtained from ESCHERICHIA COLI.	E coli Proteins
D039002	Prophages	Genomes of temperate BACTERIOPHAGES integrated into the DNA of their bacterial host cell. The prophages can be duplicated for many cell generations until some stimulus induces its activation and virulence.	Prophage