Patrycja Zalas-Więcek

Comparison of detection methods for extended-spectrum beta-lactamases in Escherichia coli strains.

INTRODUCTION Detection of extended-spectrum beta-lactamases (ESBLs) could be a major challenge for microbiologists--the difficulties arise mainly from the phenotypic differences among strains. MATERIALS AND METHODS Evaluation of ESBLs was performed on 42 strains of E. coli by: 1) DDST on MHA, 2) DDST on MHA with cloxacillin, 3) CT on MHA, according to CLSI, 4) CT on MHA with cloxacillin, 5) Etest ESBL (AB Biodisk), 6) CHROMagarTM ESBL (GRASO), 7) ChromID® ESBL (bioMérieux), and 8) automatic system VITEK2 ESBL test (bioMérieux). RESULT Positive results were obtained for 20 strains using method 1, for 18 strains using method 2, 17 by method 3, 14 by method 4, 11 by method 5, 39 by method 6, 40 by method 7, and 15 by method 8. Using Etest ESBL 6.0 non-determinable results were obtained. The most consistent results were obtained when comparing the results of method 3 with results of method 2 (97.6%), and comparing the results obtained using methods 3 and 8 (95.2%). CONCLUSIONS Based on our study we conclude that the chromogenic media can only be used as a screening method for the detection of ESBLs in E. coli rods. Etest is less useful compared to other phenotype methods, due to the impossibility of obtaining results for all the tested strains. Adding cloxacillin to MHA does not increase the frequency of detection of ESBLs in E. coli strains. DDST seems to be the most reliable among phenotypic methods for the detection of ESBLs in E. coli rods.

Diversity of extended-spectrum beta-lactamase-producing Escherichia coli rods

INTRODUCTION The aim of the study was to evaluate genetic relatedness and antimicrobial susceptibility of extended-spectrum beta-lactamase-producing E. coli strains isolated from patients hospitalized in the University Hospital in Bydgoszcz (Poland). MATERIAL AND METHODS The study included 33 extended-spectrum beta-lactamase-producing E. coli strains isolated from 31 patients. The chromosomal DNA was extracted from the strains and separated by pulsed-field gel electrophoresis. Antimicrobial susceptibility testing was performed by the agar dilution method and carried out according to the European Committee on Antimicrobial Susceptibility Testing recommendations. RESULTS According to the pulsed-field gel electrophoresis results, 32 distinct pulsotypes were revealed. Based on Molecular Analyst Fingerprinting software analysis, the studied isolates were divided into four subgroups: 6 (18.2%) isolates showing similarity greater than 90% (group A); 19 (57.6%) showing 80-90% similarity (group B); 7 (21.2%) showing 70-79% similarity (group C); and one isolate with less than 70% similarity (group D). Among E. coli isolates showing similarity greater than 90%, four antimicrobial patterns were noted. Among the isolates showing 80-90% similarity, 18 antimicrobial patterns were observed. E. coli isolates showing 70-79% similarity presented 6 antimicrobial patterns. CONCLUSIONS Our results show a high degree of genetic diversity of extended-spectrum beta-lactamase-producing E. coli isolates. However, based on a similarity of ≥80%, almost 75% of E. coli isolates were clonally related. Although it is difficult to identify definitive transmission events based on the recovery of indistinguishable pulsed-field gel electrophoresis types alone, we speculate that extended-spectrum beta-lactamase-producing E. coli strains may have disseminated throughout the hospital.