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Epidemiology

Outbreak of Infection Caused by Enterobacter cloacae Producing the Novel VEB-3 Beta-Lactamase in China

Xiaofei Jiang, Yuxing Ni, Yanqun Jiang, Feiyi Yuan, Lizhong Han, Ming Li, Hong Liu, Li Yang, Yuan Lu
Xiaofei Jiang
1Center of Laboratory Medicine of Huashan Hospital, Shanghai Medical College, Fudan University
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Yuxing Ni
2Department of Clinical Microbiology, Rijin Hospital, Shanghai Second Medical University
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Yanqun Jiang
3Department of Clinical Laboratory of the Sixth People's Hospital of Shanghai, Shanghai, People's Republic of China
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Feiyi Yuan
1Center of Laboratory Medicine of Huashan Hospital, Shanghai Medical College, Fudan University
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Lizhong Han
2Department of Clinical Microbiology, Rijin Hospital, Shanghai Second Medical University
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Ming Li
1Center of Laboratory Medicine of Huashan Hospital, Shanghai Medical College, Fudan University
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Hong Liu
1Center of Laboratory Medicine of Huashan Hospital, Shanghai Medical College, Fudan University
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Li Yang
2Department of Clinical Microbiology, Rijin Hospital, Shanghai Second Medical University
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Yuan Lu
1Center of Laboratory Medicine of Huashan Hospital, Shanghai Medical College, Fudan University
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  • For correspondence: yuanlu@hsh.stn.sh.cn
DOI: 10.1128/JCM.43.2.826-831.2005
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  • FIG. 1.
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    FIG. 1.

    Comparison of the amino acid sequences of the VEB-1, VEB-1a, VEB-1b, VEB-2, and VEB-3 beta-lactamases from P. aeruginosa JES from Thailand, P. aeruginosa KU-1 and KU-2 from Kuwait, P. aeruginosa 16 from Thailand, and E. cloacae 39 from China. Dashes indicate identical amino acids. The arrow indicates the position of the putative cleavage site of the leader peptide. Numbering is according to Ambler.

  • FIG. 2.
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    FIG. 2.

    Schematic representation of the three integrons found in E. cloacae 39, 42, and 57 and the structure of the blaVEB-3 gene cassette. Gene cassettes are shown as boxes, with arrows indicating the orientation of transcription and black circles indicating the 59-base element. The 5′ conserved segment contains the intI1 gene, which encodes integrase. The ereA2 gene was interrupted by IS1247, which are indicated by filled and empty triangles, respectively.

  • FIG. 3.
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    FIG. 3.

    (a) Plasmid DNA profiles of clinical isolates E. cloacae 39 and 57; (b) Southern blots hybridized with an intI1-specific probe; (c) Southern blots hybridized with a blaVEB-1-like specific probe. Lanes A, V517 plasmid size marker; lanes M, 1-kb DNA marker; lanes 1, plasmid DNA profiles of E. cloacae 39; lanes 2, plasmid DNA profiles of E. cloacae 57; lanes 3, chromosomal DNA of E. cloacae 39; lanes 4, chromosomal DNA of E. cloacae 57.

  • FIG. 4.
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    FIG. 4.

    (a) rep-PCR profiles of E. cloacae clinical isolates with primer ERIC2; (b) rep-PCR profiles of E. cloacae clinical isolates with primer REP. Lanes M, DNA molecular size markers; lanes A to G, rep-PCR profiles of seven distinct types of E. cloacae clinical isolates, respectively. Subscript numerals: 1, ERIC2 type; 2, REP PCR type.

Tables

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  • TABLE 1.

    Sequences of primers used for detection of bla genes or genotyping of strains

    ESBL or regionPrimerSequencePositionGenBank accession no. or reference
    TEMTEM-A5′-ATA AAA TTC TTG AAG AC-3′1-17 X54604.1
    TEM-B5′-TTA CCA ATG CTT AAT CA-3′1075-1059
    SHVSHV-FOR5′-TGGTTATGCGTTATATTCGCC-3′69-89 X98100.1
    SHV-REV5′-GCTTAGCGTTGCCAGTGCT-3′936-918
    PRE-1PER-1-FOR5′-AATTTGGGCTTAGGGCAGAA-3′278-306 Z21957
    PER-1-REV5′-ATGAATGTCATTATAAAAGC-3′1211-1192
    VEB-1VEB-1-FOR5′-CGACTTCCATTTCCCGATGC-3′226-245 AF220758
    VEB1-REV5′-GGACTCTGCAACAAATACGC-3′868-849
    CTX-M-3M13U5′-GGTTAAAAAATCACTGCGTC-3′65-84 X92506
    M13L5′-TTGGTGACGATTTTAGCCGC-3′928-909
    CTX-M-9M9U5′-ATGGTGACAAAGAGAGTGCA-3′1-20 AF252621.2
    M9L5′-CCC TTC GGC GAT GAT TCT C-3′870-852
    CTX-M-2M25U5′-ATGATGACTCAGAGCATTCG-3′304-323 AJ416343.1
    M25L5′-TGGGTTACGATTTTCGCCGC-3′1169-1150
    VEBcasVEBcas-F5′-GTTAGCGGTAATTTAACCAGATAG-3′11-34 AY027870.1
    VEBcas-B5′-CGGTTTGGGCTATGGGCAG-3′1081-1063
    Rep-PCRERIC25′-AAGTAAGTGACTGGGGTGAGCG-3′ 22
    REP 2-15′-ICG ICT TAT CIG GCC TAC-3′ 2
    VEBINV15′-CAGTTTGAGCATTTGAATACAC-3′ 16
    VEBINV25′-AGC GTA TTT GTT GCA GAG TCC-3′
    Variable region of integron5-CS5′-GGC ATC CAA GCA GCA AGC-3′1163-1180 AF205943
    3-CS5′-AAG CAG ACT TGA CCT GAT-3′9231-9214
    VEB cloneVEBc-R5′-CCGGGATCCATGAAAATCGTAAAAAGGATATTATT-3′ AF220758
    VEBc-L5′-CCCAAGCTTTTATTTATTCAAATAGTAATTCCACG-3
  • TABLE 2.

    MICs of selected antibiotics for representative E. cloacae clinical isolates

    E. cloacae isolateEtest MIC (μg/ml)a
    IPMFEPAKCAZCTXPTcCIPTZ-TZLFOXCT-CTL
    720.38312483240.13>32/>4>256>16/>1
    640.254848>25620.25>32/>4>256>16/>1
    50.25>2562>256>256>256>32>32/>4>256>16/>1
    570.2524>256>256>256>256>32>32/>4>256>16/>1
    1360.2531.512>25660.75>32/>4>256>16/>1
    420.5128>256>256>256>256>32>32/>4>256>16/>1
    1500.38>256>256>256>256>256>32>32/>448>16/>1
    1080.25>256>256>256>256>256>32>32/>4>256>16/>1
    390.75>256>256>256>256>256>32>32/>4>256>16/>1
    1030.250.7549632960.16>32/>4>256>16/>1
    2120.52>2563648>32>32/>4>256>16/>1
    1420.2532>256>256>2560.06>32/>4>256>16/>1
    • ↵ a Abbreviations: IPM, imipenem; FEP, cefepime; AK, amikacin; CAZ, ceftazidime; CTX, cefotaxime; PTc, cefoperazone-sulbactam; CIP, ciprofloxacin; TZ-TZL, ceftazidime-ceftazidime-clavulanic acid; FOX, cefoxitin; CT-CTL, cefotaxime-cefotaxime-clavulanic acid.

  • TABLE 3.

    Genotyping of rep-PCR products and beta-lactamase genes of 27 ceftazidime-resistant or cefotaxime-resistant E. cloacae clinical isolates and correlation with phenotypic ESBL tests

    TypeIsolate(s)Result for bla gene for:Result by NCCLS K-Ba method
    TEM-1CTX-M-3CTX-M-9aVEB-1SHV-5a
    A5, 25−−+−−+b
    B64−−+−−+
    12, 150+−+−−−
    33++−−++
    39, 40, 42, 71++−+−−
    236, 57, 108++−+−−
    77, 88++−+−−
    157, 205, 189+−−+−−
    17, 224++−−−−
    C72++−−−+
    D103−−−−−−
    E136+−+−−−
    216−−++−−
    F140, 212−−−−−−
    G142+−−−−−
    • ↵ a K-B, Kirby-Bauer disk diffusion.

    • ↵ b Isolate 5 was negative.

  • TABLE 4.

    MICs of beta-lactams for recombinant bacteria E. coli EV3 and E. coli ET

    E. coli strainEtest MIC (μg/ml)a
    IPMFEPCAZCTXPTcCAZ-TZLFOXCTX-CTLAM
    EV30.163>256240.38>32/0.251.58/0.25>256
    ET0.1250.0320.470.160.380.5/0.0941.50.5/0.0942
    • ↵ a Abbreviations: IPM, imipenem; FEP, cefepime; CAZ, ceftazidime; CTX, cefotaxime; PTc, cefoperazone-sulbactam; TZL, ceftazidime-clavulanic acid; FOX, cefoxitin; CTL, cefotaxime-clavulanic acid, AM, ampicillin.

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Outbreak of Infection Caused by Enterobacter cloacae Producing the Novel VEB-3 Beta-Lactamase in China
Xiaofei Jiang, Yuxing Ni, Yanqun Jiang, Feiyi Yuan, Lizhong Han, Ming Li, Hong Liu, Li Yang, Yuan Lu
Journal of Clinical Microbiology Feb 2005, 43 (2) 826-831; DOI: 10.1128/JCM.43.2.826-831.2005

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Outbreak of Infection Caused by Enterobacter cloacae Producing the Novel VEB-3 Beta-Lactamase in China
Xiaofei Jiang, Yuxing Ni, Yanqun Jiang, Feiyi Yuan, Lizhong Han, Ming Li, Hong Liu, Li Yang, Yuan Lu
Journal of Clinical Microbiology Feb 2005, 43 (2) 826-831; DOI: 10.1128/JCM.43.2.826-831.2005
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KEYWORDS

Cross Infection
Disease Outbreaks
Enterobacter cloacae
Enterobacteriaceae Infections
beta-lactamases

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