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Mechanisms of Resistance

Increase in Quinolone Resistance in aHaemophilus influenzae Strain Isolated from a Patient with Recurrent Respiratory Infections Treated with Ofloxacin

Jordi Vila, Joaquim Ruiz, Ferran Sanchez, Ferran Navarro, Beatriz Mirelis, M. Teresa Jimenez de Anta, Guillem Prats
Jordi Vila
Departament de Microbiologia, Institut de Investigació Biomédica August Pi i Sunyer, Hospital Clı́nic, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, and
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Joaquim Ruiz
Departament de Microbiologia, Institut de Investigació Biomédica August Pi i Sunyer, Hospital Clı́nic, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, and
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Ferran Sanchez
Departament de Genètica i Microbiologia, Universitat Autonoma de Barcelona, Hospital de Sant Pau, 08025 Barcelona, Spain
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Ferran Navarro
Departament de Genètica i Microbiologia, Universitat Autonoma de Barcelona, Hospital de Sant Pau, 08025 Barcelona, Spain
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Beatriz Mirelis
Departament de Genètica i Microbiologia, Universitat Autonoma de Barcelona, Hospital de Sant Pau, 08025 Barcelona, Spain
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M. Teresa Jimenez de Anta
Departament de Microbiologia, Institut de Investigació Biomédica August Pi i Sunyer, Hospital Clı́nic, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, and
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Guillem Prats
Departament de Genètica i Microbiologia, Universitat Autonoma de Barcelona, Hospital de Sant Pau, 08025 Barcelona, Spain
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DOI: 10.1128/AAC.43.1.161
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ABSTRACT

The increase in the level of quinolone resistance ofHaemophilus influenzae clinical isolates during ofloxacin therapy of a patient with recurrent respiratory infections was investigated. The first isolate (MIC of ciprofloxacin of 2 μg/ml) and the second isolate (MIC of 32 μg/ml) belonged to the same clone, as shown by pulsed-field gel electrophoresis, and the increase in the resistance level was associated with a substitution in Ser-84 to Arg in the ParC protein. These results emphasize the potential risk of development of quinolone-resistant H. influenzae during fluoroquinolone therapy in patients with recurrent respiratory infection.

Haemophilus influenzae is a frequent cause of upper and lower respiratory tract infections (9). The new fluoroquinolones have been widely used as therapy for respiratory tract infections and have shown good activity against H. influenzae (3). Resistance to quinolones in H. influenzae is mainly due to chromosomal mutations in the gyrA and parC genes, which encode the A subunits of the DNA gyrase and topoisomerase IV, respectively (2, 7), similar to those found in other bacterial species (11, 12). Although ciprofloxacin-resistantH. influenzae strains have been isolated (2, 4), the development of quinolone resistance in H. influenzaefrom patients with chronic lung disease has been infrequently reported, and the mechanism of quinolone resistance acquisition has not been investigated (1).

We studied the increase in the level of quinolone resistance of anH. influenzae clinical isolate during ofloxacin therapy in a patient with recurrent respiratory infections. The patient was a 59-year-old female with severe bronchiectasis and recurrent respiratory infections repeatedly submitted to multiple courses of antibiotics (frequently including amoxicillin plus clavulanic acid or ciprofloxacin). In May 1997, she was admitted to the hospital for an episode of bronchial infection, respiratory failure, and severe hypoxemia. She was on ventilatory support and intravenous therapy. Sputum culture for noncapsulated H. influenzae (isolate 1) with susceptibility to ciprofloxacin (MIC, 2 μg/ml), ofloxacin (MIC, 4 μg/ml), and nalidixic acid (MIC, ≥256 μg/ml) was positive. The patient was treated with ofloxacin (200 mg/12 h orally) for 4 days and subsequently with amoxicillin plus clavulanic acid and was discharged after compensation. Four months later, she attended the outpatient clinic and a control sputum culture was positive, with two colonial morphotypes of H. influenzae being detected. One (isolate 2) had the same resistance pattern as the one isolated in May (isolate 1), while the only difference with the other (isolate 3) was that the MIC of ciprofloxacin was 32 μg/ml (isolate 3). Finally, three months later, the patient was again admitted for respiratory deterioration, and two H. influenzae isolates with resistance patterns identical to those previously recovered were found in sputum. The epidemiological relationship of these isolates was investigated by pulsed-field gel electrophoresis (PFGE), showing that the three isolates belonged to the same clone (Fig.1).

Fig. 1.
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Fig. 1.

Molecular typing of H. influenzae strains by PFGE. Lanes 1, 2, and 10 are molecular weight markers. Lanes 3, 4, and 5 are isolates 1, 2, and 3 of this study, respectively. Lanes 6, 7, 8, and 9 are different strains of H. influenzae chosen randomly.

MICs were determined by a commercial microdilution test (Emiza 2E; Sensititre Ltd., Imberhorne, United Kingdom) and for nalidixic acid, by the E-test method (AB Biodisk, Dalvagen, Sweden) performed according to the manufacturers’ instructions. In addition, for the strain with a MIC of ciprofloxacin higher than 2 μg/ml, the MIC was determined by the macrodilution broth method according to the National Committee for Clinical Laboratory Standards recommendations (10).

PCR amplification was used to amplify the quinolone resistance-determining region (QRDR) of the gyrA andparC genes, and the nucleotide sequences of these amplicons were determined. The oligonucleotide primers used to amplify the QRDR of the gyrA gene from nucleotides 17 to 816 (800 bp) (from codon 6 to 272) were 5′ AATCATCTATCACCCCTGTC 3′ and 5′ TTTTGCTTTATTTACTTGGT 3′, whereas for the amplification of the QRDR of the parC gene from nucleotides 95 to 471 (377 bp) (from codon 32 to 157) the oligonucleotide primers used were 5′ ATCGTGCGTTGCCTTTTATC 3′ and 5′ TTCAGCCAAGGTTCCATCAA 3′. The PCR program and the DNA sequencing methodology used were as described in reference11.

Nucleotide sequencing of the 800- and 377-bp amplicons obtained from the QRDR of the gyrA and parC genes, respectively, revealed several mutations leading to the amino acid substitutions shown in Table 1.

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Table 1.

Mutations in the gyrA and parCgenes of different isolates of H. influenzae

The substitution at amino acid 84 (Ser-Leu) of the GyrA protein or its equivalent in other microorganisms is the most prevalent and has been found in H. influenzae (2, 7) and in other bacteria (11, 12). Georgiou et al. (7) found that strains with MICs of 2 μg/ml showed double mutations, one in the amino acid codon Asp-88 of the gyrA gene and another in the amino acid codon Ser-84 of the parC gene. Similarly, isolates 1 and 2 (MICs of ciprofloxacin of 2 μg/ml) of our study also present a double mutation, whereas the strain for which the MIC of ciprofloxacin was 32 μg/ml showed three main substitutions, two in the GyrA protein (Ser-84 to Leu and Asp-88 to Ala) and one in the ParC protein (Ser-84 to Arg). These results are also in agreement with those found by Georgiou et al. (7). The mutation in the amino acid codon Asp-83, which generated a substitution to Asn, is apparently neutral despite the change in the charge.

Studying H. influenzae in sputum samples, Groeneveld et al. (8) found patients persistently infected with the sameH. influenzae strain for up to 23 months. Similarly, the strain described herein persisted for at least 7 months despite the treatment with amoxicillin plus clavulanic acid.

These results emphasize the potential risk of development of quinolone-resistant H. influenzae during fluoroquinolone therapy in patients with recurrent respiratory infections and confirm the role of substitutions in positions Ser-84 and Asp-88 of the GyrA protein and Ser-84 of the ParC protein in the acquisition of quinolone resistance in this microorganism.

ACKNOWLEDGMENTS

This work was supported in part by grants SAF97/0091 and FIS 97/0623 from Spain.

FOOTNOTES

    • Received 26 February 1998.
    • Returned for modification 1 July 1998.
    • Accepted 5 October 1998.
  • Copyright © 1999 American Society for Microbiology

REFERENCES

  1. 1.↵
    1. Barriere S. L.,
    2. Hindler J. A.
    Ciprofloxacin-resistant Haemophilus influenzae infection in a patient with chronic lung disease. Ann. Pharmacother. 27 1993 309 310
    OpenUrlPubMedWeb of Science
  2. 2.↵
    1. Bootsma H. J.,
    2. Troeltra A.,
    3. van Veen-Rutgers A.,
    4. Mooi F. R.,
    5. de Neeling A. J.,
    6. Overbeek B. P.
    Isolation and characterization of a ciprofloxacin-resistant isolate of Haemophilus influenzae from The Netherlands. J. Antimicrob. Chemother. 39 1997 292 293
    OpenUrlCrossRefPubMedWeb of Science
  3. 3.↵
    1. Brueggeman A. B.,
    2. Kugler K. C.,
    3. Doern G. V.
    In vitro activity of BAY 12-8039, a novel 8-methoxyquinolone, compared to activities of six fluoroquinolones against Streptococcus pneumoniae,Haemophilus influenzae, and Moraxella catarrhalis. Antimicrob. Agents Chemother. 41 1997 1594 1597
    OpenUrlAbstract/FREE Full Text
  4. 4.↵
    1. Campos J.,
    2. Román F.,
    3. Georgiou M.,
    4. Garcia C.,
    5. Gomez-Lus R.,
    6. Cantón R.,
    7. Escobar V.,
    8. Baquero F.
    Long-term persistence of ciprofloxacin-resistant Haemophilus influenzae in patients with cystic fibrosis. J. Infect. Dis. 174 1996 1345 1347
    OpenUrlCrossRefPubMedWeb of Science
  5. 5.
    1. Chamberland S.,
    2. Malouin F.,
    3. Schollaardt T.,
    4. Parr T. R. Jr.,
    5. Bryan L. E.
    Persistence of Pseudomonas aeruginosa during ciprofloxacin therapy of a cystic fibrosis patient: transient resistance to quinolones and protein F-deficiency. J. Antimicrob. Chemother. 25 1990 995 1010
    OpenUrlCrossRefPubMed
  6. 6.
    1. Corkill J. E.,
    2. Percival A.,
    3. McDonald P.,
    4. Bamber A. I.
    Detection of quinolone resistance in Haemophilus spp. J. Antimicrob. Chemother. 34 1994 841 844
    OpenUrlCrossRefPubMedWeb of Science
  7. 7.↵
    1. Georgiou M.,
    2. Muñoz R.,
    3. Román F.,
    4. Cantón R.,
    5. Gómez-Lus R.,
    6. Campos J.,
    7. de la Campa A. G.
    Ciprofloxacin-resistant Haemophilus influenzae strains possess mutations in analogous positions of GyrA and ParC. Antimicrob. Agents Chemother. 40 1996 1741 1744
    OpenUrlAbstract/FREE Full Text
  8. 8.↵
    1. Groeneveld K.,
    2. van Alphen L.,
    3. Eijk P. P.,
    4. Visschers G.,
    5. Jansen H. M.,
    6. Zanen H. C.
    Endogenous and exogenous reinfections by Haemophilus influenzae in patients with chronic obstructive pulmonary disease: the effect of antibiotic treatment of persistence. J. Infect. Dis. 161 1990 512 517
    OpenUrlCrossRefPubMedWeb of Science
  9. 9.↵
    1. Moxon E. R.
    Haemophilus influenzae Principles and practice of infectious diseases. Mandell G. L., Douglas J. E., Dolin R. 1995 2039 2045 Churchill Livingstone New York, N.Y
  10. 10.↵
    National Committee for Clinical Laboratory Standards Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically 4th ed. Approved standard M7-A4. 1997 National Committee for Clinical Laboratory Standards Wayne, Pa
  11. 11.↵
    1. Vila J.,
    2. Ruiz J.,
    3. Marco F.,
    4. Barcelo A.,
    5. Goñi P.,
    6. Giralt E.,
    7. Jimenez de Anta T.
    Association between double mutation in gyrA gene of ciprofloxacin-resistant clinical isolates of Escherichia coli and MICs. Antimicrob. Agents Chemother. 38 1994 2477 2479
    OpenUrlAbstract/FREE Full Text
  12. 12.↵
    1. Vila J.,
    2. Ruiz J.,
    3. Goñi P.,
    4. Jimenez de Anta M. T.
    Detection of mutations in parC in quinolone-resistant clinical isolates of Escherichia coli. Antimicrob. Agents Chemother. 40 1996 491 493
    OpenUrlAbstract/FREE Full Text
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Increase in Quinolone Resistance in aHaemophilus influenzae Strain Isolated from a Patient with Recurrent Respiratory Infections Treated with Ofloxacin
Jordi Vila, Joaquim Ruiz, Ferran Sanchez, Ferran Navarro, Beatriz Mirelis, M. Teresa Jimenez de Anta, Guillem Prats
Antimicrobial Agents and Chemotherapy Jan 1999, 43 (1) 161-162; DOI: 10.1128/AAC.43.1.161

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Increase in Quinolone Resistance in aHaemophilus influenzae Strain Isolated from a Patient with Recurrent Respiratory Infections Treated with Ofloxacin
Jordi Vila, Joaquim Ruiz, Ferran Sanchez, Ferran Navarro, Beatriz Mirelis, M. Teresa Jimenez de Anta, Guillem Prats
Antimicrobial Agents and Chemotherapy Jan 1999, 43 (1) 161-162; DOI: 10.1128/AAC.43.1.161
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KEYWORDS

anti-infective agents
Haemophilus Infections
Haemophilus influenzae
ofloxacin
respiratory tract infections

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