title: Rapid and specific detection of Escherichia coli clonal group A by gene-specific PCR.
abstract: PCR primers specific for the recently described antimicrobial resistance-associated Escherichia coli clonal group A (CGA), a widespread cause of drug-resistant urinary tract infections in the United States, were devised on the basis of a novel single-nucleotide polymorphism identified within the housekeeping gene fumC, i.e., C288T. In comparison with two reference PCR-based fingerprinting methods, ERIC2 PCR and random amplified polymorphic DNA (RAPD) analysis, a PCR assay incorporating the new primers provided 100% sensitivity and 100% specificity for the detection of CGA among 138 diverse clinical and reference E. coli isolates. E. coli reference (ECOR) strain 47 was shown to be a member or a close relative of CGA (by ERIC2 PCR and RAPD analysis, respectively) and yielded a positive assay result. The new CGA-specific PCR assay, which exhibited interlaboratory reproducibility and stability under various experimental conditions, should allow the rapid and specific detection of CGA by any laboratory equipped for diagnostic PCR.
authors.affiliation: Infectious Diseases (111F), Minneapolis VA Medical Center, One Veterans Dr., Minneapolis, MN 55417, USA. johns007@umn.edu
authors.forename: James R
authors.initials: JR
authors.lastname: Johnson
authors.forename: Krista
authors.initials: K
authors.lastname: Owens
authors.forename: Amee R
authors.initials: AR
authors.lastname: Manges
authors.forename: Lee W
authors.initials: LW
authors.lastname: Riley
mesh_terms: D017931:DNA Primers; D004926:Escherichia coli / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D015203:Reproducibility of Results
chemical_list: D017931:DNA Primers