Skip to main content
PMC home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1991 Dec;35(12):2574–2579. doi: 10.1128/aac.35.12.2574

Comparison of conventional susceptibility tests with direct detection of penicillin-binding protein 2a in borderline oxacillin-resistant strains of Staphylococcus aureus.

J L Gerberding 1, C Miick 1, H H Liu 1, H F Chambers 1
PMCID: PMC245433  PMID: 1810191

Abstract

Six selected strains of Staphylococcus aureus classified as borderline oxacillin-resistant, according to standard disk diffusion and microdilution susceptibility test methods, and seven methicillin-resistant and seven methicillin-susceptible control strains were examined for the presence of penicillin-binding protein 2a (PBP 2a) by fluorography and immunoblotting and for DNA hybridization with a mec-specific probe in a dot blot assay. Oxacillin agar screen tests with and without NaCl supplementation were also performed with all strains. PBP 2a was detected both by fluorography and by immunoblotting in all seven methicillin-resistant control strains and in none of the susceptible controls. PBP 2a was detected in two borderline strains. Results of agar screen tests performed without NaCl supplementation were completely concordant with susceptibility determined by PBP 2a and mec detection methods. Agar screening with NaCl supplementation was less accurate. These findings were confirmed with 20 additional borderline strains. Direct detection methods for the presence of PBP 2a or mec, the gene encoding it, allow accurate and definitive classification of borderline strains. Further efforts to develop a rapid, clinically useful, antibody detection system for PBP 2a are warranted.

Full text

PDF
2574

Images in this article

2577Image
on p.2577
2577Image
on p.2577
2577Image
on p.2577
2577Image
on p.2577

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Archer G. L., Pennell E. Detection of methicillin resistance in staphylococci by using a DNA probe. Antimicrob Agents Chemother. 1990 Sep;34(9):1720–1724. doi: 10.1128/aac.34.9.1720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BARBER M. Methicillin-resistant staphylococci. J Clin Pathol. 1961 Jul;14:385–393. doi: 10.1136/jcp.14.4.385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berger-Bächi B., Barberis-Maino L., Strässle A., Kayser F. H. FemA, a host-mediated factor essential for methicillin resistance in Staphylococcus aureus: molecular cloning and characterization. Mol Gen Genet. 1989 Oct;219(1-2):263–269. doi: 10.1007/BF00261186. [DOI] [PubMed] [Google Scholar]
  4. Brown D. F., Reynolds P. E. Intrinsic resistance to beta-lactam antibiotics in Staphylococcus aureus. FEBS Lett. 1980 Dec 29;122(2):275–278. doi: 10.1016/0014-5793(80)80455-8. [DOI] [PubMed] [Google Scholar]
  5. Chambers H. F., Archer G., Matsuhashi M. Low-level methicillin resistance in strains of Staphylococcus aureus. Antimicrob Agents Chemother. 1989 Apr;33(4):424–428. doi: 10.1128/aac.33.4.424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chambers H. F. Coagulase-negative staphylococci resistant to beta-lactam antibiotics in vivo produce penicillin-binding protein 2a. Antimicrob Agents Chemother. 1987 Dec;31(12):1919–1924. doi: 10.1128/aac.31.12.1919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Coudron P. E., Jones D. L., Dalton H. P., Archer G. L. Evaluation of laboratory tests for detection of methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis. J Clin Microbiol. 1986 Nov;24(5):764–769. doi: 10.1128/jcm.24.5.764-769.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fluit A. C., Box A. T., Verhoef J. A probe for the detection of methicillin-resistant Staphylococcus aureus. Eur J Clin Microbiol Infect Dis. 1990 Aug;9(8):605–608. doi: 10.1007/BF01967216. [DOI] [PubMed] [Google Scholar]
  9. Georgopapadakou N. H., Smith S. A., Bonner D. P. Penicillin-binding proteins in a Staphylococcus aureus strain resistant to specific beta-lactam antibiotics. Antimicrob Agents Chemother. 1982 Jul;22(1):172–175. doi: 10.1128/aac.22.1.172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hartman B. J., Tomasz A. Expression of methicillin resistance in heterogeneous strains of Staphylococcus aureus. Antimicrob Agents Chemother. 1986 Jan;29(1):85–92. doi: 10.1128/aac.29.1.85. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hartman B. J., Tomasz A. Low-affinity penicillin-binding protein associated with beta-lactam resistance in Staphylococcus aureus. J Bacteriol. 1984 May;158(2):513–516. doi: 10.1128/jb.158.2.513-516.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Liu H., Buescher G., Lewis N., Snyder S., Jungkind D. Detection of borderline oxacillin-resistant Staphylococcus aureus and differentiation from methicillin-resistant strains. Eur J Clin Microbiol Infect Dis. 1990 Oct;9(10):717–724. doi: 10.1007/BF02184683. [DOI] [PubMed] [Google Scholar]
  13. Massanari R. M., Pfaller M. A., Wakefield D. S., Hammons G. T., McNutt L. A., Woolson R. F., Helms C. M. Implications of acquired oxacillin resistance in the management and control of Staphylococcus aureus infections. J Infect Dis. 1988 Oct;158(4):702–709. doi: 10.1093/infdis/158.4.702. [DOI] [PubMed] [Google Scholar]
  14. McDougal L. K., Thornsberry C. The role of beta-lactamase in staphylococcal resistance to penicillinase-resistant penicillins and cephalosporins. J Clin Microbiol. 1986 May;23(5):832–839. doi: 10.1128/jcm.23.5.832-839.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Montanari M. P., Tonin E., Biavasco F., Varaldo P. E. Further characterization of borderline methicillin-resistant Staphylococcus aureus and analysis of penicillin-binding proteins. Antimicrob Agents Chemother. 1990 May;34(5):911–913. doi: 10.1128/aac.34.5.911. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Murakami K., Tomasz A. Involvement of multiple genetic determinants in high-level methicillin resistance in Staphylococcus aureus. J Bacteriol. 1989 Feb;171(2):874–879. doi: 10.1128/jb.171.2.874-879.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Thauvin-Eliopoulos C., Rice L. B., Eliopoulos G. M., Moellering R. C., Jr Efficacy of oxacillin and ampicillin-sulbactam combination in experimental endocarditis caused by beta-lactamase-hyperproducing Staphylococcus aureus. Antimicrob Agents Chemother. 1990 May;34(5):728–732. doi: 10.1128/aac.34.5.728. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Thornsberry C., McDougal L. K. Successful use of broth microdilution in susceptibility tests for methicillin-resistant (heteroresistant) staphylococci. J Clin Microbiol. 1983 Nov;18(5):1084–1091. doi: 10.1128/jcm.18.5.1084-1091.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tomasz A., Drugeon H. B., de Lencastre H. M., Jabes D., McDougall L., Bille J. New mechanism for methicillin resistance in Staphylococcus aureus: clinical isolates that lack the PBP 2a gene and contain normal penicillin-binding proteins with modified penicillin-binding capacity. Antimicrob Agents Chemother. 1989 Nov;33(11):1869–1874. doi: 10.1128/aac.33.11.1869. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. de Lencastre H., Sá Figueiredo A. M., Urban C., Rahal J., Tomasz A. Multiple mechanisms of methicillin resistance and improved methods for detection in clinical isolates of Staphylococcus aureus. Antimicrob Agents Chemother. 1991 Apr;35(4):632–639. doi: 10.1128/aac.35.4.632. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES