Among the clinical isolates from February to July, 2001 at Showa University Hospital, 5 of 205
E. coli (2.4%), 3 of 93
K. pneumoniae (3.2%) and 2 of 52
K. oxytoca (3.8%) were extended spectrum β-lactamase (ESBLs) -producing strains identified by a combination of minimal inhibitory concentration (MIC) detection and clavulanic acid inhibition tests. After Spe I digestion of the genome DNA from the above ESBL-producing isolates, pulsed field gel electrophoretic analysis revealed that 2 of the 5
E. coli, 2 of the 3
K. pneumoniaeand 2 of the 2
K.oxytocastrains had identical genome types, respectively. Despite the identical genome type, however, MIC patterns of the 2
E. coli strains and of the 2
K.oxytocawere quite different. By PCR analysis, the presence of the TEM type β-lactamase gene was confirmed in the 2 strains of
E. coliand 1 of 2 strains of
K. pneumoniae, although the 2 stains of
K. pneumoniashowed a similar MIC pattern and an identical genome type. Sequence analysis showed that the sequences of the TEM type β-lactamase gene from the three positive strains were not identical. The nucleotides of the gene had mutations in the
E. coliand
K. pneumoniae strains, and the mutations would result in amino acid substitutions. These mutations seem to be unique because they are different from the well-known mutations of the gene reported previously. The above results indicate that detection of the MIC pattern or analysis of the resistant gene is not enough to trace the routes of infection. Identification of the genome type may also be essential for epidemiological analysis of such nosocomial infections as ESBL-producing Gram-negative rods.
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