title: Determination of flagellar types by PCR-RFLP analysis of enteropathogenic Escherichia coli (EPEC) and Shiga toxin-producing E. coli (STEC) strains isolated from animals in São Paulo, Brazil.
abstract: This study evaluated the polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP) analysis of fliC for typing flagella antigen (H) of Shiga toxin-producing Escherichia coli (STEC) and enteropathogenic E. coli (EPEC) strains isolated from different animals. The molecular typing of the H type was efficient in the determination of 93 (85%) strains. Two nonmotile (H-) E. coli strains showed a PCR-RFLP electrophoretic profile that did not match known H type patterns. The fliC nucleotide sequence of strains B2N and 4a revealed a nucleotide substitution at the restriction site and a nucleotide insertion that generated a stop codon, respectively. The results of this study showed that PCR-RFLP analysis of fliC is faster, less laborious and as efficient for the determination of H type E. coli isolated from animals, compared to serotyping and that it is useful in determining H type in nonmotile strains and strains expressing non-reactive H antigens. Moreover, the fliC sequence of strain B2N suggests that we could have found a new flagellin antigen type.
authors.affiliation: Departamento de Microbiologia, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: Claudia de Oliveira
authors.initials: Cde O
authors.lastname: Ayala
authors.forename: Ana Carolina
authors.initials: AC
authors.lastname: Ramos Moreno
authors.forename: Marina Baquerizo
authors.initials: MB
authors.lastname: Martinez
authors.forename: Ylanna Kelner
authors.initials: YK
authors.lastname: Burgos
authors.forename: Antonio Fernando
authors.initials: AF
authors.lastname: Pestana de Castro
authors.forename: Silvia Yumi
authors.initials: SY
authors.lastname: Bando
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics; D001483:Base Sequence; D001938:Brazil; D002417:Cattle; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D004285:Dogs; D054308:Enteropathogenic Escherichia coli / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D029968:Escherichia coli Proteins / Q000235:genetics*; D005408:Flagellin; D000882:Haplorhini; D008969:Molecular Sequence Data; D058889:Molecular Typing / Q000379:methods / Q000662:veterinary*; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA / Q000662:veterinary; D012756:Sheep; D054323:Shiga-Toxigenic Escherichia coli / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D000942:Antigens, Bacterial; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; C516095:FliC protein, E coli; C022863:H antigen; D005408:Flagellin