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Mechanisms of Resistance

Clinical Strains of Pseudomonas aeruginosa Overproducing MexAB-OprM and MexXY Efflux Pumps Simultaneously

Catherine Llanes, Didier Hocquet, Christelle Vogne, Dounia Benali-Baitich, Catherine Neuwirth, Patrick Plésiat
Catherine Llanes
1Laboratoire de Bactériologie, Hôpital Jean Minjoz, Besançon
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  • For correspondence: catherine.llanes-barakat@univ-fcomte.fr
Didier Hocquet
1Laboratoire de Bactériologie, Hôpital Jean Minjoz, Besançon
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Christelle Vogne
1Laboratoire de Bactériologie, Hôpital Jean Minjoz, Besançon
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Dounia Benali-Baitich
1Laboratoire de Bactériologie, Hôpital Jean Minjoz, Besançon
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Catherine Neuwirth
2Laboratoire de Bactériologie Médicale, Hôpital du Bocage, Dijon, France
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Patrick Plésiat
1Laboratoire de Bactériologie, Hôpital Jean Minjoz, Besançon
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DOI: 10.1128/AAC.48.5.1797-1802.2004
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Tables

  • TABLE 1.

    Primers used for DNA amplification

    Primer5′ → 3′ nucleotide sequenceReference
    nalC1TCA ACC CTA ACG AGA AAC GCTThis study
    nalC2TCC ACC TCA CCG AAC TGCThis study
    mexA-1CGA CCA GGC CGT GAG CAA GCA GCDumas et al. (submitted)
    mexA-2GGA GAC CTT CGC CGC GTT GTC GCDumas et al. (submitted)
    mexX-1TGA AGG CGG CCC TGG ACA TCA GCDumas et al. (submitted)
    mexX-2GAT CTG CTC GAC GCG GGT CAG CGDumas et al. (submitted)
    PA3720-1TCG CCC TGG TCT ATC CGC CGC TCThis study
    PA3720-2CCG CTC AGC AGT GCC TTC GCC ATThis study
    rpsL-1GCA ACT ATC AAC CAG CTG GTGDumas et al. (submitted)
    rpsL-2GCT GTG CTC TTG CAG GTT GTGDumas et al. (submitted)
  • TABLE 2.

    Susceptibilities of P. aeruginosa strains to antimicrobial agents

    StrainGenotypeAminoglycoside- modifying enzymeMICa (μg/ml)
    nalagrTicCazFepAtmAmkTobAprForCip
    PAO11622440.516160.12
    Mut-Gr1 agrZ 1624481 64 64 0.5
    PT629 nalB 64 44 16 40.516160.25
    ATM4 nalB agrZ 64 4 16 16 81NDND 1
    WL22 nalB agrZ More than 2b 128 8 16 32 128 128 64 128 64
    WL24 nalD agrW ANT(2′′)-I 128 8 16 32 16 64 64 128 64
    1113 nalD agrZ 64 4 8 16 8132 128 0.5
    1217 nalD agrZ More than 2 64 4 16 16 64 32 128 128 64
    1237 nalB agrZ AAC(6′)-II 64 4 8 16 16 128 64 128 16
    1250 nalC agrZ 64 4 8 16 811632 0.5
    1562 nalD agrZ More than 2 64 4 16 16 128 64 128 128 32
    1727 nalB nalC agrW 128 8 16 32 32 2 64 128 2
    1738 nalC agrW 64 2 8 16 8132 128 0.5
    2085 nalB agrZ AAC(3)-VI322 8 16 16 4 64 128 8
    2151 nalB agrZ 322 8 16 81 64 128 32
    2172 nalC agrZ 64 4 8 16 81 64 128 1
    • ↵ a Tic, ticarcillin; Caz, ceftazidime; Fep, cefepime; Atm, aztreonam; Amk, amikacin; Tob, tobramycin; Apr, apramycin; For, fortimicin; Cip, ciprofloxacin. Values in boldface are at least fourfold higher than that for PAO1. ND, not determined.

    • ↵ b Complex susceptibility profile suggesting the production of two or more different modifying enzymes.

  • TABLE 3.

    Genetic analysis of MexAB-OprM- and MexXY-overproducing strains of P. aeruginosa

    StrainNucleotide sequencing dataaMean gene expressionb
    mexZcmexRdPA3721eVicinity of PA3721mexXmexAPA3720
    PAO11.01.01.0
    PT629 (nalB)ND4 bp deleted at posi- tion 3630.63.61.71
    ATM4 (nalB agrZ) CAG→TAG (C307T) GCA→CCA (A66P)4826.50.11
    WL224844, 12671, 153G−73→A, A+13→T1689.22.9
    WL2471, 20922.05.10.2
    1113 GCC→-CC (A35−), ATG→ AT-(M34−)12671, 209T−2→C, C inserted (−25)29.64.45.5
    1217144, GAG→-AG (E157−)7198.56.00.5
    123748, CAG→TAG (Q10Z)44, 12671, 153, 209, TCA→GCA (S46A)A+13→T2727.73.7
    1250GGT→G--(G46−), GCG→- CG (A47−)500 bp deleted31217.9147
    1562144711298.51.7
    17277-bp insertion between 355 and 36171, GAT→GAA (D76E)30034.89.9
    173871, 209, CTG→CCG (L61P)82.211.7224
    20854844, 12671, 153T−2→C2048.75.8
    2151GTC→GCC (V44A)44, 12671, 153, 209T−2→C, A+13→T1102.66.7
    2172GGT→AGT (G46S)71, ATG→ACG (M151T)95.46.1194
    • ↵ a Nucleotide differences compared with the PAO1 genome (www.pseudomonas.com ).

    • ↵ b Relative to PAO1; mean values from two independent experiments.

    • ↵ c 48, substitution GTC→GCC (V48→A); 144, GCG→GTG (V144→A).

    • ↵ d 44, substitution AAG→ATG (K44→M); 126, GTG→GAG (V126→E).

    • ↵ e 71, substitution GGG→GAG (G71→E); 153, GAG→CAG (E153→Q); 209, AGC→CGC (S209→R).

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Clinical Strains of Pseudomonas aeruginosa Overproducing MexAB-OprM and MexXY Efflux Pumps Simultaneously
Catherine Llanes, Didier Hocquet, Christelle Vogne, Dounia Benali-Baitich, Catherine Neuwirth, Patrick Plésiat
Antimicrobial Agents and Chemotherapy Apr 2004, 48 (5) 1797-1802; DOI: 10.1128/AAC.48.5.1797-1802.2004

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Clinical Strains of Pseudomonas aeruginosa Overproducing MexAB-OprM and MexXY Efflux Pumps Simultaneously
Catherine Llanes, Didier Hocquet, Christelle Vogne, Dounia Benali-Baitich, Catherine Neuwirth, Patrick Plésiat
Antimicrobial Agents and Chemotherapy Apr 2004, 48 (5) 1797-1802; DOI: 10.1128/AAC.48.5.1797-1802.2004
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KEYWORDS

Bacterial Outer Membrane Proteins
Bacterial Proteins
Membrane Transport Proteins
Pseudomonas Infections
Pseudomonas aeruginosa

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