ABSTRACT
OBJECTIVE: To characterize the mechanism of glycopeptide resistance and to determine the genetic relatedness among strains by pulsed-field gel electrophoresis (PFGE) in vancomycin-resistant Enterococcus faecium from Argentina. MATERIALS AND METHODS: A total of 189 vancomycin-resistant single-patient isolates of Enterococcus faecium recovered between January 1997 and December 2000 from 30 hospitals in Argentina were studied. Minimum inhibitory concentrations were determined by the agar dilution method and van genes were detected by PCR. PFGE was used for molecular typing. RESULTS: All isolates except three (vanB) were of genotype vanA. For 189 vancomycin-resistant Enterococcus faecium, SmaI-PFGE indicated 35 clonal types. Most of the isolates (56%) belonged to the same clonal type 1, which was present in 19 hospitals and dominant in 17. CONCLUSIONS: The emergence of vancomycin-resistant Enterococcus faecium in Argentina seems to be related to the intra- and inter-hospital dissemination of an epidemic clone carrying the vanA element.
Subject(s)
Cross Infection/microbiology , Enterococcus faecium/genetics , Gram-Positive Bacterial Infections/microbiology , Vancomycin Resistance/genetics , Argentina/epidemiology , Bacterial Proteins/genetics , Carbon-Oxygen Ligases/genetics , Cross Infection/epidemiology , Electrophoresis, Gel, Pulsed-Field/methods , Enterococcus faecium/drug effects , Enterococcus faecium/isolation & purification , Gram-Positive Bacterial Infections/epidemiology , Humans , Microbial Sensitivity Tests , Molecular EpidemiologyABSTRACT
The gene bla(CARB-9) was located in the Vibrio cholerae super-integron, but in a different location relative to bla(CARB-7). CARB-9 (pI 5.2) conferred beta-lactam MICs four to eight times lower than those conferred by CARB-7, differing at Ambler's positions V97I, L124F, and T228K. Comparison of the genetic environments of all reported bla(CARB) genes indicated that the CARB enzymes constitute a family of cassette-encoded beta-lactamases.
Subject(s)
Penicillinase/genetics , Vibrio cholerae/enzymology , Vibrio cholerae/genetics , beta-Lactamases/genetics , Amino Acid Sequence , Argentina , Molecular Sequence Data , Penicillinase/classification , Repetitive Sequences, Amino Acid , beta-Lactamases/classification , beta-Lactamases/metabolismABSTRACT
The method of normalized resistance interpretation (NRI), uses the high-zone side of the susceptible peak in a zone diameter histogram as an internal calibrator to construct the real standard distribution of susceptible isolates even in the presence of resistant isolates. NRI parameters were optimized using control strain histograms from microbiology laboratories in Stockholm, Argentina, and the Philippines. A moving average based on four-zone values was slightly better than based on two-zone average values. The optimal peak adjustment from the switch position of the moving average was 1.0 for two-zone averages and 2.5 for four-zone averages. A comparison between true means and NRI-calculated means showed a highly significant correlation (R(2)=0.963). Coefficients of variation (CV), comparing the CV of the true distribution of control strain test results with the NRI calculated distribution, identified two types of aberrant histograms. NRI calculations on clinical isolates of Escherichia coli and Staphylococcus aureus from selected laboratories showed a good agreement between the local resistance interpretations with the NRI calculated levels. One type of deviation was most marked with cephalothin histograms for E. coli isolates where the regular zone breakpoints used cut through the population of susceptible strains. With proper markers for required quality of disc test results, the NRI method might be a valuable tool for both resistance surveillance and for quality control of the disc diffusion method.
