Sequence of the Escherichia coli O26 O antigen gene cluster and identification of O26 specific genes

D'Souza, Jocelyne M.; Wang, Lei; Reeves, Peter R.

doi:10.1016/s0378-1119(02)00876-4

Cited by 59 publications

(66 citation statements)

References 24 publications

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“…Furthermore, the sequences obtained after a mutagenesis assay showed a high degree of identity with genes involved in LPS biosynthesis pathways. The fnl-1 gene is one of the three genes that seem to be responsible for the synthesis of N-acetyl-L-fucosamine, one of the sugars composing the O26 O antigen (7). The wbjB gene from Pasteurella multocida and Pseudomonas aeruginosa and the rfb gene cluster of L. interrogans are also involved in the O-antigen biosynthesis pathway (6,16).…”

Section: Discussionmentioning

confidence: 99%

“…The wbjB gene from Pasteurella multocida and Pseudomonas aeruginosa and the rfb gene cluster of L. interrogans are also involved in the O-antigen biosynthesis pathway (6,16). The wbuA and wbuB genes belong to the O-antigen gene cluster from the O26:K60: H311b E. coli strain and code for a putative rhamnosyl transferase and a putative L-fucosamine transferase, respectively (7).…”

Section: Discussionmentioning

confidence: 99%

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2F3 Monoclonal Antibody Recognizes the O26 O-Antigen Moiety of the Lipopolysaccharide of Enterohemorrhagic Escherichia coli Strain 4276

Szalo

Taminiau

Goffaux

et al. 2004

Clin Vaccine Immunol

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Enterohemorrhagic Escherichia coli (EHEC) and enteropathogenic E. coli (EPEC) organisms are groups of pathogenic strains whose infections are characterized by a typical lesion of enterocyte attachment and effacement. They are involved in enteric diseases both in humans and in animals, and EHEC strains can be responsible for hemolytic uremic syndrome in humans. Previously, it was shown that the 2F3 monoclonal antibody (MAb) is specific for the O26 EHEC and EPEC strains (P. Enterohemorrhagic Escherichia coli (EHEC) strains are an important class of pathogens for humans and animals. The term EHEC denotes E. coli strains that cause attaching and effacing lesions on epithelial cells, express Shiga toxin-encoding genes, possess a 60-MDa virulence plasmid, and produce hemorrhagic colitis and/or hemolytic uremic syndrome (17,18).KerrBesides those of the O157:H7 and O157:H Ϫ serotypes, EHEC strains of serogroup O26 are the most common strains found in human infections (4, 17). In addition, O26 EHEC strains are an important cause of enteritis in calves (12). One of the most important steps in the pathogenic mechanism of enteric bacteria is the initial adhesion of the bacteria to the intestinal wall. Several stages of colonization have been recognized in attaching and effacing E. coli pathogenesis, some of which await characterization. Some level of host specificity is shown by EHEC strains, but the role in host specificity of the recognized virulence factors, such as Shiga toxin 1 (Stx1) and/or Stx2 and the intimin protein (a protein that is encoded by the eae gene and necessary for the production of the attaching and effacing lesion), is unknown (8, 19). The bovine host specificity shown by O26 EHEC and enteropathogenic E. coli (EPEC) strains could be based on the production of a colonization factor, such as an adhesin, that is specific for cattle; such a colonization factor has been demonstrated to be the basis of the host specificity of classical human EPEC strains. Such an adhesin is likely to be a specific immunogen exposed at the bacterial cell surface (3,13,15,17,23,24).A monoclonal antibody (MAb), 2F3, was derived against a surface antigen extracted from the bovine O26 EHEC strain 4276; it tested positive in an enzyme-linked immunosorbent assay (ELISA) with all of the O26 and half of the O111 bovine and human EHEC and EPEC strains (9). Recent reexamination of six of the O111 EHEC and EPEC strains that tested positive revealed a contamination by O26 EHEC or EPEC strains. Purification of the strains in the mixed culture demonstrated that the O111 strains were 2F3 negative and the O26 strains were 2F3 positive (J. McCappin and H. Ball, unpublished data).Since the 2F3 MAb is specific for the O26 EHEC and EPEC strains, it may not only represent a diagnostic tool of these confirmed pathogens but also recognize a factor involved in the production of the attaching and effacing lesion. In view of these results, this study was initiated (i) to characterize the antigen recognized by the 2F3 MAb and its genetic determinant an...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

2F3 Monoclonal Antibody Recognizes the O26 O-Antigen Moiety of the Lipopolysaccharide of Enterohemorrhagic Escherichia coli Strain 4276

Szalo

Taminiau

Goffaux

et al. 2004

Clin Vaccine Immunol

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“…The sequences of sugar transferase genes and O-unitprocessing genes are normally specific to a particular O antigen. Specific PCR methods based on O-antigen-specific genes have been proposed for molecular typing of many E. coli and Shigella O serogroups (21,25,26,28,54,(57)(58)(59).…”

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confidence: 99%

“…ORFs 11, 12, and 13 showed 81, 57, and 70% identity to FnlA (WbjB), FnlB (WbjC), and FnlC (WbjD), respectively, of the Pseudomonas aeruginosa O11 O-antigen gene cluster (GenBank entries AAF72954, AAF72955, and AAF72956) (19); 79, 43, and 50% identity to FnlA (Cap5E), FnlB (Cap5F), and FnlC (Cap5G), respectively, of the Staphylococcus aureus type 5 capsule gene cluster (GenBank entries AAC46088, AAC46089, and AAC46090); and 88, 71, and 89% identity to FnlA, FnlB, and FnlC, respectively, of the E. coli O26 O-antigen gene cluster (21). FnlA, FnlB, and FnlC are enzymes of the UDP-L-FucNAc biosyn- (GenBank accession number AAG06538) and shared the motif SGGLDSS with homologues of WbpG.…”

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confidence: 99%

Structural and Genetic Characterization of Enterohemorrhagic Escherichia coli O145 O Antigen and Development of an O145 Serogroup-Specific PCR Assay

et al. 2005

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Enterohemorrhagic Escherichia coli O145 strains are emerging as causes of hemorrhagic colitis and hemolytic uremic syndrome. In this study, we present the structure of the E. coli O145 O antigen and the sequence of its gene cluster. The O145 antigen has repeat units containing three monosaccharide residues: 2-acetamido-2-deoxy-D-glucose (GlcNAc), 2-acetamidoylamino-2,6-dideoxy-L-galactose, and N-acetylneuraminic acid. It is very closely related to Salmonella enterica serovar Touera and S. enterica subsp. arizonae O21 antigen. The E. coli O145 gene cluster is located between the JUMPStart sequence and the gnd gene and consists of 15 open reading frames. Putative genes for the synthesis of the O-antigen constituents, for sugar transferase, and for O-antigen processing were annotated based on sequence similarities and the presence of conserved regions. The putative genes located in the E. coli O145 O-antigen gene cluster accounted for all functions expected for synthesis of the structure. An E. coli O145 serogroup-specific PCR assay based on the genes wzx and wzy was also developed by screening E. coli and Shigella isolates of different serotypes.

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“…Our group, among others, has been working on DNA-based O-serotyping methods in Shigella and E. coli with an objective of developing a DNA-based typing scheme. Recently, a number of reports have shown that using PCR assays based on O-antigen specific genes can distinguish particular Shigella and E. coli O-serogroups (Wang et al 1998;Wang et al 1998;Wang et al 2001a;D'Souza et al 2002;Perelle et al 2002;DebRoy et al 2003;Fratamico et al 2003;Feng et al 2004aFeng et al , 2004bFeng et al , 2004cTao et al 2004). As an alternative to the conventional serotyping assay, which is time consuming, subject to serological cross-reactions, and inaccurate because of the presence of smooth and rough forms of LPS produced by bacteria, the PCR method is accurate, sensitive, and rapid.…”

Section: Introductionmentioning

confidence: 99%

Molecular analysis ofShigella boydiiO1 O-antigen gene cluster and its PCR typing

Jiang¹,

Wang²,

Liú³

et al. 2005

Can. J. Microbiol.

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Shigella is an important human pathogen and is closely related to Escherichia coli. O-antigen is the most variable part of the lipopolysaccharide on the cell surface of Gram-negative bacteria and plays an important role in pathogenicity. The O-antigen gene cluster of S. boydii O1 was sequenced. The putative genes encoding enzymes for rhamnose synthesis, transferases, O-unit flippase, and O-unit polymerase were identified on the basis of homology. The O-antigen gene clusters of S. boydii O1 and E. coli O149, which share the same O-antigen form, were found to have the same genes and organization by adjacent gene PCR assay. Two genes specific for S. boydii O1 and E. coli O149 were identified by PCR screening against E. coli-and Shigella-type strains of the 186 known O-antigen forms and 39 E. coli clinical isolates. A PCR sensitivity of 10 3 to 10 4 CFU/mL overnight culture of S. boydii O1 and E. coli O149 was obtained. S. boydii O1 and E. coli O149 were differentiated by PCR using lacZ-and cadA-based primers.

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Sequence of the Escherichia coli O26 O antigen gene cluster and identification of O26 specific genes

Cited by 59 publications

References 24 publications

2F3 Monoclonal Antibody Recognizes the O26 O-Antigen Moiety of the Lipopolysaccharide of Enterohemorrhagic Escherichia coli Strain 4276

2F3 Monoclonal Antibody Recognizes the O26 O-Antigen Moiety of the Lipopolysaccharide of Enterohemorrhagic Escherichia coli Strain 4276

Structural and Genetic Characterization of Enterohemorrhagic Escherichia coli O145 O Antigen and Development of an O145 Serogroup-Specific PCR Assay

Molecular analysis ofShigella boydiiO1 O-antigen gene cluster and its PCR typing

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