title: Incidence and mechanisms of resistance to the combination of amoxicillin and clavulanic acid in Escherichia coli.
abstract: Among Escherichia coli organisms isolated at St. Thomas's Hospital during the years 1990 to 1994, the frequency of resistance to amoxicillin-clavulanic acid (tested by disk diffusion in a ratio of 2:1) remained constant at about 5% of patient isolates (10 to 15% of the 41 to 45% that were amoxicillin resistant). Mechanisms of increased resistance were determined for 72 consecutively collected such amoxicillin-clavulanic acid-resistant isolates. MICs of the combination were 16-8 micrograms/ml for 51 (71%) of these and > or = 32-16 micrograms/ml for the remainder. The predominant mechanism was hyperproduction of enzymes isoelectrically cofocusing with TEM-1 (beta-lactamase activities, > 200 nmol of nitrocefin hydrolyzed per min per mg of protein) which was found in 44 isolates (61%); two isolates produced smaller amounts (approximately 150 nmol/min/mg) of such enzymes, and two isolates hyperproduced enzymes cofocusing with TEM-2. Eleven isolates produced enzymes cofocusing with OXA-1 beta-lactamase, which has previously been associated with resistance to amoxicillin-clavulanic acid. Ten isolates produced increased amounts of chromosomal beta-lactamase, and four of these additionally produced TEM-1 or TEM-2. Three isolates produced inhibitor-resistant TEM-group enzymes. In one of the enzymes (pI, 5.4), the amino acid sequence change was Met-67-->Val, and thus the enzyme is identical to TEM-34. Another (pI, 5.4) had the substitution Met-67-->Ile and is identical to IRT-I67, which we propose now be given the designation TEM-40. The third (pI, 5.2) had the substitution Arg-241-->Thr; this enzyme has not been reported previously and should be called TEM-41. The rarity and diversity of inhibitor-resistant TEM-group enzymes suggest that they are the result of spontaneous mutations that have not yet spread.
authors.affiliation: Department of Microbiology, UMDS, London, United Kingdom.
authors.forename: P
authors.initials: P
authors.lastname: Stapleton
authors.forename: P J
authors.initials: PJ
authors.lastname: Wu
authors.forename: A
authors.initials: A
authors.lastname: King
authors.forename: K
authors.initials: K
authors.lastname: Shannon
authors.forename: G
authors.initials: G
authors.lastname: French
authors.forename: I
authors.initials: I
authors.lastname: Phillips
mesh_terms: D000658:Amoxicillin / Q000494:pharmacology; D019980:Amoxicillin-Potassium Clavulanate Combination; D000668:Ampicillin Resistance; D001483:Base Sequence; D002969:Clavulanic Acids / Q000494:pharmacology; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D004352:Drug Resistance, Microbial; D004359:Drug Therapy, Combination / Q000494:pharmacology*; D004926:Escherichia coli / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics; D007525:Isoelectric Focusing; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D010406:Penicillins / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D001618:beta-Lactamases / Q000378:metabolism
chemical_list: D002969:Clavulanic Acids; D004269:DNA, Bacterial; D010406:Penicillins; D019980:Amoxicillin-Potassium Clavulanate Combination; D000658:Amoxicillin; D001618:beta-Lactamases