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Mechanisms of Resistance

Prevalence of SHV-12 among Clinical Isolates ofKlebsiella pneumoniae Producing Extended-Spectrum β-Lactamases and Identification of a Novel AmpC Enzyme (CMY-8) in Southern Taiwan

Jing-Jou Yan, Shiou-Mei Wu, Shu-Huei Tsai, Jiunn-Jong Wu, Ih-Jen Su
Jing-Jou Yan
Department of Pathology, National Cheng Kung University Medical Center, and
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Shiou-Mei Wu
Department of Medical Technology, National Cheng Kung University Medical College, Tainan, Taiwan
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Shu-Huei Tsai
Department of Pathology, National Cheng Kung University Medical Center, and
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Jiunn-Jong Wu
Department of Medical Technology, National Cheng Kung University Medical College, Tainan, Taiwan
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Ih-Jen Su
Department of Pathology, National Cheng Kung University Medical Center, and
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DOI: 10.1128/AAC.44.6.1438-1442.2000
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    Fig. 1.

    Complete nucleotide sequence and predicted amino acid sequence of the blaCMY-8 β-lactamase gene. The primers for amplification of the gene were synthesized according to the nucleotide sequence of CMY-1 (4, 14) and are double underlined. DNA sequencing was performed with a sequencing primer (underlined) and the PCR primers as well. Arrows indicate the directions of DNA sequencing. The stop codon is indicated with three asterisks. The β-lactamase active site S-V-S-K, the conserved triad K-T-G, and the class C typical motif Y-X-N are underlined.

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    Fig. 2.

    Alignment of deduced amino acid sequences of CMY-8 with those of CMY-1 and MOX-1. Identical amino acids are marked with dots. The stop codon is indicated with an asterisk. The β-lactamase active site S-V-S-K, the conserved triad K-T-G, and the class C typical motif Y-X-N are shown in boldface type.

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    Fig. 3.

    PFGE of XbaI-digested genomic DNAs from 19 ESBL-producing K. pneumoniae isolates. Lanes 1, 12, 13, and 23, bacteriophage lambda DNA concatemers (GibcoBRL, Gaithersburg, Md.) which served as molecular size marker; lanes 2 to 11 and 14 to 22, isolates V293, D155, B657, C106, O574, R549, O787, W142, W580, T384, X386, C097, Q381, T923, T386, R436, T986, B262, and T255, respectively. Isolate S276 is not shown here. All ESBL producers except for three isolates carrying CMY-8 (W142, W580, and T384) had different PFGE patterns.

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  • Table 1.

    MICs for K. pneumoniae strains with different resistance phenotypes and their transconjugantsa

    Resistance phenotype groupNo. of clinical isolatesMIC (μg/ml) of drug for clinical isolatesNo. of transconjugantsMIC (μg/ml) of drug for transconjugants
    AMPAMCCAZCTXFOXIPMAMPAMCCAZCTXFOXIPM
    I14>2564–8≥2563–163–160.125–0.2511>2564–8≥2566–162–60.125–0.25
    II2>2564–82–24128–2568–160.1252>2564–82–3128–2568–160.125
    III3>25616 96>256>2560.25–0.53>2562432–64>256>2560.25–0.5
    IV1>25612>256 48>2560.51>25612>256 8>2560.125
    • ↵a MICs were determined by the E test. Abbreviations: AMP, ampicillin; AMC, amoxicillin-clavulanic acid combination, CAZ, ceftazidime; CTX, cefotaxime; FOX, cefoxitin; IPM, imipenem.

  • Table 2.

    Results of IEF, genotyping of β-lactamases, plasmid analysis, and PFGE

    Resistance phenotype group and strainClinical isolateTransconjugantc
    pI(s)SHVTEMAmpCPlasmid profilePFGE patternpISHVAmpCApprox plasmid size (kb)
    Group I
     X3868.2, 7.65AI8.25100
     C0978.2, 7.6, 5.451BII8.25100
     Q3818.2, 7.6, 5.451CIII8.2570
     T923a8.2, 7.6, 5.451DIV
     T3868.2, 7.6, 5.4121EV8.21270
     R4368.2, 7.6, 5.4121EVI8.212100
     S2768.2, 7.6, 5.4121FVII8.212100
     T9868.2, 7.6, 5.4121GVIII8.21270
     B262a8.2, 7.6, 5.4121HIX
     T255a8.2, 7.6, 5.4121IX
     V2938.2, 7.6, 5.4121GXI8.21270
     D1558.2, 7.612JXII8.21240
     B6578.2, 7.612KXIII8.21275
     C1068.2, 7.612JXIV8.21240
    Group II
     O5748.3, 7.6, 5.4111LXV8.395
     R5498.3, 7.6, 5.4111MXVI8.3125
    Group III
     W1428.25, 7.6, 5.4111CMY-8NXVII8.25CMY-825
     W5808.25, 7.6, 5.4111CMY-8NXVII8.25CMY-825
     T3848.25, 7.6, 5.4111CMY-8NXVII8.25CMY-825
    Group IV
     O7878.2, 7.6, 5.4121UDbOXVIII8.212UD150
    • ↵a Conjugation experiments with strains T923, B262, and T255 were not successful.

    • ↵b UD, undetermined.

    • ↵c All transconjugates lacked TEM.

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Prevalence of SHV-12 among Clinical Isolates ofKlebsiella pneumoniae Producing Extended-Spectrum β-Lactamases and Identification of a Novel AmpC Enzyme (CMY-8) in Southern Taiwan
Jing-Jou Yan, Shiou-Mei Wu, Shu-Huei Tsai, Jiunn-Jong Wu, Ih-Jen Su
Antimicrobial Agents and Chemotherapy Jun 2000, 44 (6) 1438-1442; DOI: 10.1128/AAC.44.6.1438-1442.2000

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Prevalence of SHV-12 among Clinical Isolates ofKlebsiella pneumoniae Producing Extended-Spectrum β-Lactamases and Identification of a Novel AmpC Enzyme (CMY-8) in Southern Taiwan
Jing-Jou Yan, Shiou-Mei Wu, Shu-Huei Tsai, Jiunn-Jong Wu, Ih-Jen Su
Antimicrobial Agents and Chemotherapy Jun 2000, 44 (6) 1438-1442; DOI: 10.1128/AAC.44.6.1438-1442.2000
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KEYWORDS

Bacterial Proteins
Klebsiella Infections
Klebsiella pneumoniae
beta-lactamases

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