Prevalence of the type I and type II DHFR genes in trimethoprim-resistant urinary isolates of Escherichia coli from Greece.
Trimethoprim resistance in 64 Escherichia coli urinary isolates from five hospitals in Greece was studied. Of the 40 isolates exhibiting transferable high-level resistance (MIC > 1024 mg/L), 21 hybridized with a specific probe for dihydrofolate reductase (DHFR) I, 13 with a probe for DHFR II, and one with a probe for DHFR V. Eleven isolates hybridized with a probe for transposon Tn7. Among the 17 isolates with non-transferable high-level resistance, seven hybridized with the probe for DHFR I, three with the probe for DHFR II, and eight were Tn7-positive. None of the seven isolates with low-level resistance (MIC 4-1024 mg/L) reacted with the probes used. Of the 28 isolates positive for DHFR I, 12 (43%) failed to hybridize with the Tn7 probe. Conversely, three isolates hybridized with the Tn7 probe, but not with the probe for DHFR I. Colony hybridization experiments showed that all but three transconjugants reacted similarly to their respective parent strains. The plasmids coding for trimethoprim-resistant DHFRs were found to differ on the basis of restriction enzyme analysis. These findings suggest that trimethoprim resistance among E. coli urinary isolates in Greece is mediated predominantly by heterogeneous transferable plasmids encoding either DHFR I or DHFR II. The dissociation between DHFR I and Tn7, together with the high incidence of trimethoprim-resistant isolates which did not hybridize with the probes for the common DHFR I or II types, indicates the continued evolution of trimethoprim resistance determinants.
Tsakris
A
A
Department of Bacteriology, Hellenic Pasteur Institute, Athens, Greece.
Johnson
A P
AP
Legakis
N J
NJ
Tzouvelekis
L S
LS
D001437:Bacteriuria / Q000382:microbiology*; D003227:Conjugation, Genetic; D015342:DNA Probes; D004262:DNA Restriction Enzymes; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D006115:Greece; D006801:Humans; D008826:Microbial Sensitivity Tests; D009693:Nucleic Acid Hybridization; D010957:Plasmids / Q000235:genetics; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics*; D014296:Trimethoprim Resistance / Q000235:genetics*
D015342:DNA Probes; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D013762:Tetrahydrofolate Dehydrogenase; D004262:DNA Restriction Enzymes