Detection and characterization of the fimA gene of Escherichia coli O157:H7.
An expected 850-bp DNA fragment containing fimA, the structural gene for type 1 fimbriae, and flanking sequences was amplified from 39 (of 46) pathogenic and commensal strains of Escherichia coli using the polymerase chain reaction (PCR). Restriction fragment length polymorphism (RFLP) analysis of the amplified products showed 13 HinP1 and four Sau961 restriction profiles among these 39 E. coli strains, revealing the polymorphic nature of this allele. A unique RFLP pattern was shared by E. coli O157:H7, O157:H- and a few O55 serotype strains. DNA sequence analysis of the fimA region demonstrated that E. coli O157:H7 strain 933 and O157:H- strain E32511 contained identical DNA sequences that were distinct from other E. coli strains, especially a 16-bp sequence 5' to fimA that was conspicuously absent only in E. coli O157 strains. Exploiting these differences, a PCR assay was developed that amplifies a 936-bp fragment from all E. coli O157:H7 strains examined to date. This PCR assay offers a simple, rapid, and reliable means to detect E. coli strains of the O157:H7 serotype.
Li
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Molecular Biology Branch, U.S. Food and Drug Administration, Washington, DC 20204, USA.
Koch
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Cebula
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D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics*; D001483:Base Sequence; D019453:Escherichia coli O157 / Q000235:genetics*; D036781:Fimbriae Proteins; D005819:Genetic Markers; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D015183:Restriction Mapping; D017421:Sequence Analysis; D017384:Sequence Deletion; D012689:Sequence Homology, Nucleic Acid
D001426:Bacterial Proteins; D005819:Genetic Markers; C063055:fimbrillin; D036781:Fimbriae Proteins