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Epidemiology

Multifocal Detection of Multidrug-Resistant Pseudomonas aeruginosa Producing the PER-1 Extended-Spectrum β-Lactamase in Northern Italy

Laura Pagani, Elisabetta Mantengoli, Roberta Migliavacca, Elisabetta Nucleo, Simona Pollini, Melissa Spalla, Rossana Daturi, Egidio Romero, Gian Maria Rossolini
Laura Pagani
1Dipartimento di Scienze Morfologiche, Eidologiche e Cliniche, Sezione di Microbiologia, Università di Pavia
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Elisabetta Mantengoli
2Dipartimento di Biologia Molecolare, Sezione di Microbiologia, Università di Siena, Siena, Italy
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Roberta Migliavacca
1Dipartimento di Scienze Morfologiche, Eidologiche e Cliniche, Sezione di Microbiologia, Università di Pavia
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Elisabetta Nucleo
1Dipartimento di Scienze Morfologiche, Eidologiche e Cliniche, Sezione di Microbiologia, Università di Pavia
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Simona Pollini
2Dipartimento di Biologia Molecolare, Sezione di Microbiologia, Università di Siena, Siena, Italy
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Melissa Spalla
3Servizio di Analisi Microbiologiche IRCCSS Matteo, Pavia
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Rossana Daturi
3Servizio di Analisi Microbiologiche IRCCSS Matteo, Pavia
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Egidio Romero
1Dipartimento di Scienze Morfologiche, Eidologiche e Cliniche, Sezione di Microbiologia, Università di Pavia
3Servizio di Analisi Microbiologiche IRCCSS Matteo, Pavia
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Gian Maria Rossolini
2Dipartimento di Biologia Molecolare, Sezione di Microbiologia, Università di Siena, Siena, Italy
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  • For correspondence: rossolini@unisi.it
DOI: 10.1128/JCM.42.6.2523-2529.2004
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  • FIG. 1.
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    FIG. 1.

    Map of northern Italy showing the locations of the six hospitals from which the P. aeruginosa isolates investigated in this study were isolated. P.A.T., Pio Albergo Trivulzio Hospital; Nig., Niguarda Hospital; S.M., San Matteo Hospital. For each hospital, the total number of isolates and the number of PER-1-positive isolates are reported. The location of the hospital in Varese, from which PER-1-positive P. aeruginosa isolates were previously detected (12), is also shown (shaded in gray).

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    FIG. 2.

    PFGE profiles of genomic DNAs of PER-1-positive P. aeruginosa isolates after digestion with SpeI. The profile of isolate Pa6NI, which was identical to that of Pa34SM, and that of isolate Pa105TR, which was different from that of Pa144TR by three bands, are not shown. DNA size standards are reported on the left.

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    FIG. 3.

    Agarose gel electrophoresis of undigested genomic DNAs of the nine epidemiologically unrelated PER-1-positive P. aeruginosa isolates investigated in this study (top) and results of Southern blot analysis of the same samples with a blaPER-1 probe (bottom). DNA size standards are reported on the left.

  • FIG. 4.
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    FIG. 4.

    Results of Southern blot analysis of genomic DNAs of the nine epidemiologically unrelated PER-1-positive P. aeruginosa isolates investigated in this study with a blaPER-1 probe after digestion with EcoRI (A) or EcoRV (B). DNA size standards are reported on the left.

  • FIG. 5.
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    FIG. 5.

    Structure of SmaI-EcoRI genomic fragment containing the blaPER-1 gene from P. aeruginosa Ps101PAT and comparison with the region containing blaPER-1 from P. aeruginosa RNL-1 (16) (EMBL/GenBank database entry Z21957) and the region containing blaPER-2 in plasmid pMVP-5 from S. enterica serovar Typhymurium JMC (1) (EMBL/GenBank database entry X93314). The sequence from Ps101PAT is shown in capital letters. The start and stop codons of the blaPER genes are underlined. The start codon of the open reading frame, encoding a transposase-like protein whose translation is shown below the nucleotide sequence, is also underlined. The other sequences are shown in lowercase letters, except when they are different from that of Ps101PAT; in the region upstream of the blaPER genes, “##” indicates that the sequence completely diverges beyond that point. A vertical arrow indicates the point of insertion of an extra 115-bp sequence in the region upstream of blaPER-2.

Tables

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  • TABLE 1.

    Sources, antimicrobial susceptibilities, IEF profiles, and PFGE profiles of the nine epidemiologically unrelated PER-1-producing P. aeruginosa clinical isolates investigated in this study

    HospitalIsolateDateWardaSpecimenb typeMIC of antibiotic (μg/ml)cpIS by IEFdPFGE pattern
    CZFPATPIPTIPMECPGMTONEAK
    San Matteo, PaviaPa34SMNovember 1997HAEMCutaneous swab>32>32>323232>16>16>4>16>16>32>645.3, 8.0A
    Pa50SMMarch 1998PAED-SUUrine>32>32>32>128>128>16>16>4>16>16>32325.3, 8.3C
    Pa66SMApril 1998ICUCutaneous swab>32>32>32>128>128<4>16>4>16>16>32>645.3, 8.0D
    TreviglioPa105TRSeptember 1995ICUBAW>32>32>326416≤4>16>4>16>16>32325.3, 8.0F0
    Pa144TRSeptember 1998ICUBAW>32>32>32>12816>16>16>4>16>16>32>645.3, 8.0F1
    Pio Albergo Trivulzio, MilanPs101PATAugust 1999LCTFUrine>32>32>32>128>128>16>16>4>16>16>32>645.3, 6.9, 8.2E
    Niguarda, MilanPa6NIFebruary 1999BMTPharyngeal swab>32>32>323216>16>16>4>16>16>32>645.3, 8.0A
    Pa40NI00February 2000HAEMPharyngeal swab>32>32>3216≤8≤4>16≤1>16>16>32>645.3, 8.4B
    SondaloPa1309SODecember 1999NSUSputum>32>32>323216>16>16>4>16>16>32>645.3, 8.0G
    • ↵ a HAEM, hematology; PAED-SU, pediatric surgery; LCTF, long-term care facility; BMT, bone marrow transplantation unit; NSU, neurosurgery.

    • ↵ b BAW, bronchoalveolar washing.

    • ↵ c CZ, ceftazidime; FP, cefepime; AT, aztreonam; PI, piperacillin; PT, piperacillin-tazobactam (tazobactam at a fixed concentration of 4 μg/ml); IP, imipenem; ME, meropenem; CP, ciprofloxacin; GM, gentamicin; TO, tobramycin; NE, netilmicin; AK, amikacin.

    • ↵ d pls of β-lactamase activities in crude cell extracts, as detected by analytical IEF. In the bioassay, the pI 5.3 enzyme always exhibited activity against cefotaxime, ceftazidime, cefepime, and aztreonam; the pI 6.9 enzyme of isolate Ps101PAT exhibited activity against ceftazidime, cefepime, and aztreonam; and the pI 8.0 to 8.4 enzymes did not exhibit ESBL activity.

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Multifocal Detection of Multidrug-Resistant Pseudomonas aeruginosa Producing the PER-1 Extended-Spectrum β-Lactamase in Northern Italy
Laura Pagani, Elisabetta Mantengoli, Roberta Migliavacca, Elisabetta Nucleo, Simona Pollini, Melissa Spalla, Rossana Daturi, Egidio Romero, Gian Maria Rossolini
Journal of Clinical Microbiology Jun 2004, 42 (6) 2523-2529; DOI: 10.1128/JCM.42.6.2523-2529.2004

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Multifocal Detection of Multidrug-Resistant Pseudomonas aeruginosa Producing the PER-1 Extended-Spectrum β-Lactamase in Northern Italy
Laura Pagani, Elisabetta Mantengoli, Roberta Migliavacca, Elisabetta Nucleo, Simona Pollini, Melissa Spalla, Rossana Daturi, Egidio Romero, Gian Maria Rossolini
Journal of Clinical Microbiology Jun 2004, 42 (6) 2523-2529; DOI: 10.1128/JCM.42.6.2523-2529.2004
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KEYWORDS

Drug Resistance, Multiple, Bacterial
Pseudomonas aeruginosa
beta-lactamases

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