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Use of a DNA hybridization method to verify results of screening for methicillin resistance in Staphylococci

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Abstract

Tests were performed by the disk diffusion method, agar dilution method and the E test to determine the susceptibility to methicillin and oxacillin of clinical isolates and control strains ofStaphylococcus aureus (n=106) and coagulase-negative species (n=131). Results were compared with those of a dot blot DNA hybridization test, in which themecA gene was detected using an oligonucleotide probe selected from themecA gene. Among theStaphylococcus aureus strains themecA gene was found in all but two strains inhibited by ≥8 mg/l of methicillin and all but two strains inhibited by ≥4 mg/l of oxacillin. A disk test using either 1 µg oxacillin or 10 µg methicillin and a tentative resistance breakpoint of ≤10 mm gave the best agreement with the hybridization test. For coagulase-negative staphylococci 34 of 35 strains inhibited by ≥8 mg/l methicillin hybridized with the probe as well as 58 of 82 strains inhibited by 1–4 mg/l; 93 of 97 strains inhibited by ≥0.5 mg/l oxacillin were also positive in the probe test. Using the 1 µg oxacillin disk and a resistance breakpoint of ≤ 10 mm good agreement was obtained between results of the disk diffusion and DNA hybridization tests. It is suggested that this genotypic method for detection of methicillin resistance is used as a reference method for routine methods.

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References

  1. Cookson B, Phillips I: Methicillin-resistant staphylococci. Journal of Applied Bacteriology 1990, Supplement: 55–70.

  2. National Committee for Clinical Laboratory Standards: Performance standards for antimicrobial disk susceptibility testing. Approved standard M2-A4, fourth edition. NCCLS, Villanova, PA, 1990.

    Google Scholar 

  3. Lencastre H de, Sa Figueiredo AM, Urban C, Rahal J, Tomasz A: Multiple mechanisms of methicillin resistance and improved methods for detection in clinical isolates ofStaphylococcus aureus. Antimicrobial Agents and Chemotherapy 1991, 35: 632–639.

    PubMed  Google Scholar 

  4. Coudron PE, Jones DL, Dalton HP, Archer GL: Evaluation of laboratory tests for detection of methicillin-resistantStaphylococcus aureus andStaphylococcus epidermidis. Journal of Clinical Microbiology 1986, 24: 764–769.

    PubMed  Google Scholar 

  5. Hedin G, Hambraeus A: Screening tests for the detection of methicillin resistance inStaphylococcus epidermidis. Journal of Antimicrobial Chemotherapy 1991, 28: 681–694.

    PubMed  Google Scholar 

  6. Reynolds PE, Fuller C: Methicillin-resistant strains ofStaphyloccoccus aureus: presence of identical additional penicillin-binding protein in all strains examined. FEMS Microbiology Letters 1986, 33: 251–254.

    Article  Google Scholar 

  7. Beck WD, Berger-Bächi B, Kayser FH: Additional DNA in methicillin-resistantStaphylococcus aureus and molecular cloning ofmec-specific DNA. Journal of Bacteriology 1986, 165: 373–378.

    PubMed  Google Scholar 

  8. Ubukata K, Nonoguchi R, Song MD, Matsuhashi M, Konno M: Homology ofmecA gene in methicillin-resistantStaphylococcus haemolyticus andStaphylococcus simulans to that ofStaphylococcus aureus. Antimicrobial Agents and Chemotherapy 1990, 34: 170–172.

    PubMed  Google Scholar 

  9. Kloos WE, Lambe DW:Staphylococcus. In: Balows A, Hausler WJ, Herrmann KL, Isenberg HD, Shadomy HJ (ed): Manual of clinical microbiology. American Society for Microbiology, Washington, DC, 1991, p. 222–237.

    Google Scholar 

  10. Dornbusch K, Hallander HO, Löfquist F: Extrachromosomal control of methicillin resistance and toxin production inStaphylococcus aureus. Journal of Bacteriology 1969, 98: 351–358.

    PubMed  Google Scholar 

  11. Brown DFJ, Brown L: Evaluation of the E test, a novel method of quantifying antimicrobial activity. Journal of Antimicrobial Chemotherapy 1991, 27: 185–190.

    PubMed  Google Scholar 

  12. Song MD, Wachi M, Doi M, Ishino F, Matsuhashi M: Evolution of an inducible penicillin-target protein in methicillin-resistantStaphylococcus aureus by gene fusion. FEBS Letters 1987, 221: 167–171.

    Article  PubMed  Google Scholar 

  13. Ryffel C, Tesch W, Birch-Machin I, Reynolds PE, Barberis-Maino L, Kayser FH, Berger-Bächi B: Sequence comparison ofmecA genes isolated from methicillin-resistantStaphylococcus aureus andStaphylococcus epidermidis. Gene 1990, 94: 137–138.

    Article  PubMed  Google Scholar 

  14. Maniatis T, Fritsch EF, Sambrook J: Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, 1982.

    Google Scholar 

  15. Phillips I, Andrews AM, Bint AJ, Bridson E, Brown DFJ, Cooke EM, Greenwood D, Holt HA, King A, Spencer RC, Williams RJ, Wise R: A guide to sensitivity testing. Report of the working party on antibiotic sensitivity testing of the British Society for Antimicrobial Chemotherapy. Academic Press, London, 1991.

    Google Scholar 

  16. McDougal LK, Thornsberry C: The role of β-lactamase in staphylococcal resistance to penicillinase-resistant penicillins and cephalosporins. Antimicrobial Agents and Chemotherapy 1986, 23: 832–839.

    Google Scholar 

  17. Barry AL, Jones RN: Reliability of high-content disks and modified broth dilution tests for detecting staphylococcal resistance to the penicillinase-resistant penicillins. Journal of Clinical Microbiology 1987, 25: 1897–1901.

    PubMed  Google Scholar 

  18. Archer GL, Pennell E: Detection of methicillin resistance in staphylococci by using a DNA probe. Antimicrobial Agents and Chemotherapy 1990, 34:1720–1724.

    PubMed  Google Scholar 

  19. Courvalin P: Genotypic approach to the study of bacterial resistance to antibiotics. Antimicrobial Agents and Chemotherapy 1991, 35: 1019–1023.

    PubMed  Google Scholar 

  20. Fluit AC, Box ATA, Verhoef J: A probe for the detection of methicillin-resistantStaphylococcus aureus. European Journal of Clinical Microbiology & Infectious Diseases 1990, 9: 605–608.

    Google Scholar 

  21. Ryffel C, Kayser F, Berger-Bächi B: Correlation between regulation ofmecA transcription and expression of methicillin resistance in staphylococci. Antimicrobial Agents and Chemotherapy 1992, 36: 25–31.

    PubMed  Google Scholar 

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Olsson-Liljequist, B., Larsson, P., Ringertz, S. et al. Use of a DNA hybridization method to verify results of screening for methicillin resistance in Staphylococci. Eur. J. Clin. Microbiol. Infect. Dis. 12, 527–533 (1993). https://doi.org/10.1007/BF01970958

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