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Mechanisms of Resistance

Three Cefotaximases, CTX-M-9, CTX-M-13, and CTX-M-14, among Enterobacteriaceae in the People's Republic of China

Aroonwadee Chanawong, Fatima Hannachi M'Zali, John Heritage, Jian-Hui Xiong, Peter Michael Hawkey
Aroonwadee Chanawong
1Division of Microbiology, University of Leeds, Leeds LS2 9JT
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Fatima Hannachi M'Zali
1Division of Microbiology, University of Leeds, Leeds LS2 9JT
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John Heritage
1Division of Microbiology, University of Leeds, Leeds LS2 9JT
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Jian-Hui Xiong
2Department of Immunity and Infection, The Medical School, University of Birmingham, Birmingham B15 2TT, United Kingdom
3First Municipal People's Hospital of Guangzhou, Guangzhou 510180, People's Republic of China
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Peter Michael Hawkey
1Division of Microbiology, University of Leeds, Leeds LS2 9JT
2Department of Immunity and Infection, The Medical School, University of Birmingham, Birmingham B15 2TT, United Kingdom
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  • For correspondence: HawkeyP@heartsol.wmids.nhs.uk
DOI: 10.1128/AAC.46.3.630-637.2002
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    FIG. 1.

    Nucleotide sequence alignment of the open reading frames of CTX-M-9 36; this study), CTX-M-13 (this study), CTX-M-14 (this study), and Toho-2 (29). Dots indicate identical nucleotides. Hyphens show nucleotide deletions. Amino acids in parentheses indicate substitutions compared with the sequence of CTX-M-9. The numbering is according to Ambler et al. (1). The asterisk at the end indicates the stop codon.

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    FIG. 2.

    (a) Plasmid DNA profiles of the clinical isolates carrying blaCTX-M genes and their transconjugants; (b) Southern blots hybridized with a blaCTX-M-specific probe. Lanes A, V517 plasmid size marker; lanes B and C, E. coli strain 8 and its transconjugant (blaCTX-M-14), respectively; lanes D and E, E. coli strain 7 and its transconjugant (blaCTX-M-14), respectively; lanes F and G, K. pneumoniae strain 1 and its transconjugant (blaCTX-M-13), respectively; lanes H and I, K. pneumoniae strain 2 and its transconjugant (blaCTX-M-14), respectively; lanes J and K, E. coli strains 5 and 6, respectively (blaCTX-M-14); lanes L, E. cloacae strain 3 (blaCTX-M-14); lanes M and N, E. cloacae strain 1 and its transconjugant (blaCTX-M-9), respectively.

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  • TABLE 1.

    Oligonucleotide primers used in the study

    Gene or gene regionPrimerOligonucleotide sequence (5"→3")GenBank accession no. (reference)Position of primera
    bla CTX-M C-1AACACGGATTGACCGTCTTG D89862 (29)82-101
    C-2TTACAGCCCTTCGGCGAT D89862 981-964
    C-3GCTGGTTCTGGTGACCTATTTTACCC D89862 881-907
    C-5CCAGCAGCAGCGGAATGCAC D89862 175-156
    DownstreamC-4GTTGTCGGGAAGATACGTGA AF252622 (this study)2683-2664
    C-6CATCACGACTGTGCTGGTCATTAAAC AF252622 2860-2885
    C-7GAAATCGCACACCTGGTAATTGATTC AF252622 3035-3050
    UpstreamC-8CACTCACCTCACAAGCAACGAA AJ242809 (unpublished)902-923
    C-9AAATTGACATCCATTACGATTGA AJ242809 553-575
    C-10TGAGAATGCAAAAAGCAATGGG AJ242809 252-273
    C-11CTTCAAAAAGCATAATCAAAGCCT AJ242809 19-43
    C-12ACCATATATCGAGTCATAAAAGCTGA AJ242809 146-121
    C-13AAAAATCCCATTGCTTTTTGCATTCT AJ242809 279-254
    • ↵ a Position counted from the first nucleotide of the entire sequence, as numbered for the GenBank submission.

  • TABLE 2.

    Sources of the 15 clinical isolates used in the study

    OrganismDate collected (day/mo/yr)WardSpecimenaClinical diagnosisAntibiotic therapyb
    E. coli 113/06/97RespiratorySputumBronchopneumonia secondary to chronic bronchitisUnknown
    E. coli 215/08/97CardiologySputumBronchopneumonia secondary to chronic bronchitis and emphysemaNone
    E. coli 319/03/97PediatricNSUnknownUnknown
    E. coli 424/10/97PediatricUSNeonatal intracerebral bleedingPenicillin
    E. coli 521/10/98RespiratorySputumLow differentiation squamous cell cancer of right lung and chronic pneumoniaNone
    E. coli 623/10/98RespiratorySputumMultiple thrombi, cerebral embolus, and aspiration pneumoniaNone
    E. coli 730/10/98GastroenterologyBloodRectal carcinoma and pneumonia secondary to obstructionCeftriaxone
    E. coli 806/03/98HematologyUrineMyelodysplastic syndrome and urinary tract infectionNone
    K. pneumoniae I05/10/98NeurologySputumThrombic cerebral embolus and pneumoniaNone
    K. pneumoniae 221/10/98UrologySputumDiabetes mellitus and peripheral neuritisUnknown
    K. pneumoniae 328/10/98RespiratorySputumChronic bronchitis and chronic pulmonary emphysemaCeftriaxone
    E. cloacae 107/03/97NeurologySputumThrombic cerebral embolus and pneumoniaUnknown
    E. cloacae 224/04/97HematologySputumAcute myeloblastic leukemia with pneumoniaUnknown
    E. cloacae 307/10/98Chinese medicineSputumPneumonia secondary to Parkinson's diseaseNone
    C. freundii 18/03/97SurgerySputumLeft temporal bone defectUnknown
    • ↵ a NS, nasopharyngeal swab; US, umbilical swab.

    • ↵ b Clinical outcome data were not available.

  • TABLE 3.

    Phenotypic and genotypic characterization of the 15 clinical isolates

    Genus speciesMICa (μg/ml)pIβ-Lactamase(s) detected by PCRbPFGE patternPlasmid profile (kb)
    ATMCTXCTX+CAZCAZ+
    E. coli 112880.56445.4, 8.2SHV-12, TEM-1B∗Eco-1130, 10, 8.5
    Transconjugantc3220.5162NTdSHV-12130
    E. coli 2832≤0.252≤0.255.4, 8.1CTX-M-9, TEM-1BEco-2150, 7.0, 5.0, 4.0
    Transconjugant416≤0.251≤0.25NTCTX-M-9150, 5.0, 4.0
    E. coli 3416≤0.251≤0.255.4, 8.1CTX-M-9, TEM-1BEco-2150, 7.0, 5.0, 4.0
    Transconjugant28≤0.250.5≤0.25NTCTX-M-9150, 5.0, 4.0
    E. coli 4232NT0.5≤0.258.1CTX-M-9Eco-3150
    E. coli 5>128128NT>1280.258.1CTX-M-14, SHV-12, TEM-1BEco-4130, 3.5
    E. coli 612864NT6418.1CTX-M-14, SHV-12, TEM-1BEco-4130, 3.5
    E. coli 7432≤0.251≤0.255.4, 8.1CTX-M-14, TEM-1BFailede130
    Transconjugant416≤0.250.5≤0.25NTCTX-M-14130
    E. coli 81664≤0.2540.55.4, 8.1CTX-M-14, TEM-1B∗Eco-5130, 5.5, 4.8, 3.5, 2.5, 1.9
    Transconjugant48≤0.251≤0.25NTCTX-M-14130
    K. pneumoniae 1161281632645.4, 7.6, 8.2CTX-M-13, TEM-1B, SHV-11Kp-155, 35, 7.0
    Transconjugant832≤0.252≤0.25NTCTX-M-1335
    K. pneumoniae 2128>128≤0.253217.6, 8.1CTX-M-14, SHV-11Kp-2150, 60, 7.0
    Transconjugant432≤0.251≤0.25NTCTX-M-1460
    K. pneumoniae 31632NT8168.1CTX-M-9Kp-3150, 7.0, 2.7, 1.9
    E. cloacae 132>1280.580.58.1, 9.25CTX-M-9Ecl-185, 2.4
    Transconjugant832≤0.252≤0.25NTCTX-M-9Plasmid not found
    E. cloacae 264160.564≤0.255.4, 9.25SHV-12, TEM-1BEcl-2Plasmid not found
    Transconjugant80.5≤0.258≤0.25NTSHV-12, TEM-1BPlasmid not found
    E. cloacae 3>128128NT12848.1CTX-M-14, SHV-12Ecl-3150, 2.4
    C. freundii 1288NT128≤0.255.4, 8.2SHV-12, TEM-1BNT7.0
    • ↵ a ATM, aztreonam; CAZ, ceftazidime; CAZ+, ceftazidime plus 4 μg of clavulanic acid per ml; CTX, cefotaxime; CTX+, cefotaxime plus 4 μg of clavulanic acid per ml.

    • ↵ b The blaSHV and blaCTX-M genes were characterized by nucleotide sequence analysis, and the blaTEM gene was characterized by PCR-RFLP (2). An asterisk indicates the blaTEM gene that was confirmed by nucleotide sequence analysis.

    • ↵ c All transconjugants except those of E. cloacae 2 were E. coli UB1637 transconjugants; the E. cloacae 2 transconjugant was an E. coli UB5201 transconjugant.

    • ↵ d NT, not tested.

    • ↵ e Despite retesting, no DNA fragments were seen.

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Three Cefotaximases, CTX-M-9, CTX-M-13, and CTX-M-14, among Enterobacteriaceae in the People's Republic of China
Aroonwadee Chanawong, Fatima Hannachi M'Zali, John Heritage, Jian-Hui Xiong, Peter Michael Hawkey
Antimicrobial Agents and Chemotherapy Mar 2002, 46 (3) 630-637; DOI: 10.1128/AAC.46.3.630-637.2002

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Three Cefotaximases, CTX-M-9, CTX-M-13, and CTX-M-14, among Enterobacteriaceae in the People's Republic of China
Aroonwadee Chanawong, Fatima Hannachi M'Zali, John Heritage, Jian-Hui Xiong, Peter Michael Hawkey
Antimicrobial Agents and Chemotherapy Mar 2002, 46 (3) 630-637; DOI: 10.1128/AAC.46.3.630-637.2002
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KEYWORDS

Enterobacteriaceae
Escherichia coli Proteins
beta-lactamases

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