Skip to main content
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1992 Jun;36(6):1215–1218. doi: 10.1128/aac.36.6.1215

Identical genes confer high-level resistance to gentamicin upon Enterococcus faecalis, Enterococcus faecium, and Streptococcus agalactiae.

A Kaufhold 1, A Podbielski 1, T Horaud 1, P Ferrieri 1
PMCID: PMC190320  PMID: 1329614

Abstract

The structural gene coding for the bifunctional aminoglycoside-modifying 6'-acetyltransferase-2''-phosphotransferase (6'AAC-2''APH) enzyme was specifically amplified by the polymerase chain reaction using template DNA of clinical isolates of enterococci (both Enterococcus faecalis and Enterococcus faecium) and the single high-level gentamicin-resistant Streptococcus agalactiae strain identified thus far. The results of the present study demonstrated that the genes encoding this antibiotic resistance trait are highly homologous in these species. In dot blot hybridization assays using nonradioactively labeled oligonucleotide probes, strains with and without high-level gentamicin resistance could be discerned unequivocally. The gene segment encoding 6'-acetylating activity and the gene segment encoding 2''-phosphorylating activity were simultaneously present in all isolates exhibiting high-level gentamicin resistance.

Full text

PDF
1215

Images in this article

Selected References

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

  1. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bollet C., Gevaudan M. J., de Lamballerie X., Zandotti C., de Micco P. A simple method for the isolation of chromosomal DNA from gram positive or acid-fast bacteria. Nucleic Acids Res. 1991 Apr 25;19(8):1955–1955. doi: 10.1093/nar/19.8.1955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Buu-Hoï A., Le Bouguenec C., Horaud T. High-level chromosomal gentamicin resistance in Streptococcus agalactiae (group B). Antimicrob Agents Chemother. 1990 Jun;34(6):985–988. doi: 10.1128/aac.34.6.985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Eliopoulos G. M., Wennersten C., Zighelboim-Daum S., Reiszner E., Goldmann D., Moellering R. C., Jr High-level resistance to gentamicin in clinical isolates of Streptococcus (Enterococcus) faecium. Antimicrob Agents Chemother. 1988 Oct;32(10):1528–1532. doi: 10.1128/aac.32.10.1528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Facklam R. R., Collins M. D. Identification of Enterococcus species isolated from human infections by a conventional test scheme. J Clin Microbiol. 1989 Apr;27(4):731–734. doi: 10.1128/jcm.27.4.731-734.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ferretti J. J., Gilmore K. S., Courvalin P. Nucleotide sequence analysis of the gene specifying the bifunctional 6'-aminoglycoside acetyltransferase 2"-aminoglycoside phosphotransferase enzyme in Streptococcus faecalis and identification and cloning of gene regions specifying the two activities. J Bacteriol. 1986 Aug;167(2):631–638. doi: 10.1128/jb.167.2.631-638.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Froggatt J. W., Johnston J. L., Galetto D. W., Archer G. L. Antimicrobial resistance in nosocomial isolates of Staphylococcus haemolyticus. Antimicrob Agents Chemother. 1989 Apr;33(4):460–466. doi: 10.1128/aac.33.4.460. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hodel-Christian S. L., Murray B. E. Characterization of the gentamicin resistance transposon Tn5281 from Enterococcus faecalis and comparison to staphylococcal transposons Tn4001 and Tn4031. Antimicrob Agents Chemother. 1991 Jun;35(6):1147–1152. doi: 10.1128/aac.35.6.1147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hodel-Christian S. L., Murray B. E. Mobilization of the gentamicin resistance gene in Enterococcus faecalis. Antimicrob Agents Chemother. 1990 Jun;34(6):1278–1280. doi: 10.1128/aac.34.6.1278. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Horodniceanu T., Bougueleret L., El-Solh N., Bieth G., Delbos F. High-level, plasmid-borne resistance to gentamicin in Streptococcus faecalis subsp. zymogenes. Antimicrob Agents Chemother. 1979 Nov;16(5):686–689. doi: 10.1128/aac.16.5.686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Huang T. T., Malke H., Ferretti J. J. Heterogeneity of the streptokinase gene in group A streptococci. Infect Immun. 1989 Feb;57(2):502–506. doi: 10.1128/iai.57.2.502-506.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Huycke M. M., Spiegel C. A., Gilmore M. S. Bacteremia caused by hemolytic, high-level gentamicin-resistant Enterococcus faecalis. Antimicrob Agents Chemother. 1991 Aug;35(8):1626–1634. doi: 10.1128/aac.35.8.1626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kusukawa N., Uemori T., Asada K., Kato I. Rapid and reliable protocol for direct sequencing of material amplified by the polymerase chain reaction. Biotechniques. 1990 Jul;9(1):66-8, 70, 72. [PubMed] [Google Scholar]
  14. Kwok S., Higuchi R. Avoiding false positives with PCR. Nature. 1989 May 18;339(6221):237–238. doi: 10.1038/339237a0. [DOI] [PubMed] [Google Scholar]
  15. Mederski-Samoraj B. D., Murray B. E. High-level resistance to gentamicin in clinical isolates of enterococci. J Infect Dis. 1983 Apr;147(4):751–757. doi: 10.1093/infdis/147.4.751. [DOI] [PubMed] [Google Scholar]
  16. Patterson J. E., Zervos M. J. High-level gentamicin resistance in Enterococcus: microbiology, genetic basis, and epidemiology. Rev Infect Dis. 1990 Jul-Aug;12(4):644–652. doi: 10.1093/clinids/12.4.644. [DOI] [PubMed] [Google Scholar]
  17. Podbielski A., Kühnemund O., Lütticken R. Identification of group A type 1 streptococcal M protein gene by a non-radioactive oligonucleotide detection method. Med Microbiol Immunol. 1990;179(5):255–262. doi: 10.1007/BF00192463. [DOI] [PubMed] [Google Scholar]
  18. Rice L. B., Eliopoulos G. M., Wennersten C., Goldmann D., Jacoby G. A., Moellering R. C., Jr Chromosomally mediated beta-lactamase production and gentamicin resistance in Enterococcus faecalis. Antimicrob Agents Chemother. 1991 Feb;35(2):272–276. doi: 10.1128/aac.35.2.272. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rouch D. A., Byrne M. E., Kong Y. C., Skurray R. A. The aacA-aphD gentamicin and kanamycin resistance determinant of Tn4001 from Staphylococcus aureus: expression and nucleotide sequence analysis. J Gen Microbiol. 1987 Nov;133(11):3039–3052. doi: 10.1099/00221287-133-11-3039. [DOI] [PubMed] [Google Scholar]
  20. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ubukata K., Yamashita N., Gotoh A., Konno M. Purification and characterization of aminoglycoside-modifying enzymes from Staphylococcus aureus and Staphylococcus epidermidis. Antimicrob Agents Chemother. 1984 Jun;25(6):754–759. doi: 10.1128/aac.25.6.754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Zervos M. J., Mikesell T. S., Schaberg D. R. Heterogeneity of plasmids determining high-level resistance to gentamicin in clinical isolates of Streptococcus faecalis. Antimicrob Agents Chemother. 1986 Jul;30(1):78–81. doi: 10.1128/aac.30.1.78. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES