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PMID: 11502517 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Standardized method for in vitro antifungal susceptibility testing of Candida albicans biofilms.

Antimicrobial agents and chemotherapy ·Vol. 45 ·No. 9 ·2001-09-00 ·Pages 2475-9

Ramage G, Vande Walle K, Wickes BL, López-Ribot JL

Abstract

Candida albicans is implicated in many biomaterial-related infections. Typically, these infections are associated with biofilm formation. Cells in biofilms display phenotypic traits that are dramatically different from those of their free-floating planktonic counterparts and are notoriously resistant to antimicrobial agents. Consequently, biofilm-related infections are inherently difficult to treat and to fully eradicate with normal treatment regimens. Here, we report a rapid and highly reproducible microtiter-based colorimetric assay for the susceptibility testing of fungal biofilms, based on the measurement of metabolic activities of the sessile cells by using a formazan salt reduction assay. The assay was used for in vitro antifungal susceptibility testing of several C. albicans strains grown as biofilms against amphotericin B and fluconazole and the increased resistance of C. albicans biofilms against these antifungal agents was demonstrated. Because of its simplicity, compatibility with a widely available 96-well microplate platform, high throughput, and automation potential, we believe this assay represents a promising tool for the standardization of in vitro antifungal susceptibility testing of fungal biofilms.

MeSH Terms
Amphotericin B/pharmacology Antifungal Agents/pharmacology Biofilms/drug effects Candida albicans/drug effects Cell Count Colorimetry Fluconazole/pharmacology Humans Microbial Sensitivity Tests/standards Reproducibility of Results
Chemicals
Antifungal Agents Amphotericin B Fluconazole
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Ramage G
Department of Microbiology, Division of Infectious Diseases, The University of Texas Health Science Center at San Antonio, Texas 78245, USA.
Vande Walle K
Wickes B L
López-Ribot J L
References (37)
37 references, click to expand
  1. Candida biofilms and their susceptibility to antifungal agents.
    Methods Enzymol. 1999;310:644-56 PMID: 10547826
  2. Antibiotic susceptibility assay for Staphylococcus aureus in biofilms developed in vitro.
    J Antimicrob Chemother. 1999 Jul;44(1):43-55 PMID: 10459809
  3. Expression of the multiple antibiotic resistance operon (mar) during growth of Escherichia coli as a biofilm.
    J Appl Microbiol. 2000 Feb;88(2):243-7 PMID: 10735992
  4. Oral colonization by Candida albicans.
    Crit Rev Oral Biol Med. 1999;10(3):359-83 PMID: 10759414
  5. Riddle of biofilm resistance.
    Antimicrob Agents Chemother. 2001 Apr;45(4):999-1007 PMID: 11257008
  6. Adhesive colonization of biomaterials and antibiotic resistance.
    Biomaterials. 1987 Nov;8(6):423-6 PMID: 3427140
  7. Susceptibility of bacterial biofilms to tobramycin: role of specific growth rate and phase in the division cycle.
    J Antimicrob Chemother. 1990 Apr;25(4):585-91 PMID: 2190969
  8. The biofilm glycocalyx as a resistance factor.
    J Antimicrob Chemother. 1990 Jul;26(1):1-5 PMID: 2211430
  9. Influence of growth rate on susceptibility to antimicrobial agents: biofilms, cell cycle, dormancy, and stringent response.
    Antimicrob Agents Chemother. 1990 Oct;34(10):1865-8 PMID: 2291653
  10. Susceptibility of Pseudomonas aeruginosa and Escherichia coli biofilms towards ciprofloxacin: effect of specific growth rate.
    J Antimicrob Chemother. 1991 Feb;27(2):177-84 PMID: 1905285
  11. Bacterial resistance to antibiotics: the role of biofilms.
    Prog Drug Res. 1991;37:91-105 PMID: 1763187
  12. Innovative endpoint determination system for antifungal susceptibility testing of yeasts.
    Antimicrob Agents Chemother. 1992 Aug;36(8):1619-25 PMID: 1416843
  13. Pseudomonas aeruginosa biofilm as a diffusion barrier to piperacillin.
    Antimicrob Agents Chemother. 1992 Sep;36(9):2054-6 PMID: 1416900
  14. Production of mucoid exopolysaccharide during development of Pseudomonas aeruginosa biofilms.
    Infect Immun. 1993 Feb;61(2):777-80 PMID: 8423105
  15. Immunologic aspects of oral candidiasis.
    Oral Surg Oral Med Oral Pathol. 1994 Aug;78(2):202-10 PMID: 7936590
  16. Testing antimicrobial susceptibilities of adherent bacteria by a method that incorporates guidelines of the National Committee for Clinical Laboratory Standards.
    J Clin Microbiol. 1994 Oct;32(10):2564-8 PMID: 7814499
  17. Resistance of Candida albicans biofilms to antifungal agents in vitro.
    Antimicrob Agents Chemother. 1995 Sep;39(9):2128-31 PMID: 8540729
  18. Candida'the cancer of silastic'.
    J Laryngol Otol. 1996 Mar;110(3):240-2 PMID: 8730359
  19. Comparisons of the susceptibilities of planktonic and adherent Candida albicans to antifungal agents: a modified XTT tetrazolium assay using synchronised C. albicans cells.
    J Med Vet Mycol. 1996 Mar-Apr;34(2):149-52 PMID: 8732362
  20. Bacterial biofilms: resistance to antimicrobial agents.
    J Antimicrob Chemother. 1996 Jun;37(6):1047-50 PMID: 8836808
  21. Susceptibility testing of fungi: current status of correlation of in vitro data with clinical outcome.
    J Clin Microbiol. 1996 Mar;34(3):489-95 PMID: 8904400
  22. Adhesion of different Candida spp. to plastic: XTT formazan determinations.
    J Med Vet Mycol. 1996 Nov-Dec;34(6):407-10 PMID: 8971630
  23. Development of interpretive breakpoints for antifungal susceptibility testing: conceptual framework and analysis of in vitro-in vivo correlation data for fluconazole, itraconazole, and candida infections. Subcommittee on Antifungal Susceptibility Testing of the National Committee for Clinical Laboratory Standards.
    Clin Infect Dis. 1997 Feb;24(2):235-47 PMID: 9114154
  24. Susceptibility testing of fungi and correlation with clinical outcome.
    J Chemother. 1997 May;9 Suppl 1:19-24 PMID: 9248959
  25. The development of a small-scale biofilm model suitable for studying the effects of antibiotics on biofilms of gram-negative bacteria.
    J Antimicrob Chemother. 1997 Sep;40(3):329-34 PMID: 9338483
  26. Biofilm susceptibility to antimicrobials.
    Adv Dent Res. 1997 Apr;11(1):160-7 PMID: 9524452
  27. Cell wall and secreted proteins of Candida albicans: identification, function, and expression.
    Microbiol Mol Biol Rev. 1998 Mar;62(1):130-80 PMID: 9529890
  28. Comparison of a 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-t etrazolium hydroxide (XTT) colorimetric method with the standardized National Committee for Clinical Laboratory Standards method of testing clinical yeast isolates for susceptibility to antifungal agents.
    J Clin Microbiol. 1998 May;36(5):1450-2 PMID: 9574729
  29. Effect of growth rate on resistance of Candida albicans biofilms to antifungal agents.
    Antimicrob Agents Chemother. 1998 Aug;42(8):1900-5 PMID: 9687381
  30. Iron-limited biofilms of Candida albicans and their susceptibility to amphotericin B.
    Antimicrob Agents Chemother. 1998 Aug;42(8):2146-9 PMID: 9687427
  31. Standardization of antifungal susceptibility testing and clinical relevance.
    Med Mycol. 1998;36 Suppl 1:68-78 PMID: 9988494
  32. The Calgary Biofilm Device: new technology for rapid determination of antibiotic susceptibilities of bacterial biofilms.
    J Clin Microbiol. 1999 Jun;37(6):1771-6 PMID: 10325322
  33. Bacterial biofilms: a common cause of persistent infections.
    Science. 1999 May 21;284(5418):1318-22 PMID: 10334980
  34. Antimicrobial substances and effects on sessile bacteria.
    Zentralbl Bakteriol. 1999 Apr;289(2):165-77 PMID: 10360317
  35. Biofilms.
    Curr Opin Microbiol. 1999 Jun;2(3):270-5 PMID: 10383861
  36. In vitro development of Staphylococcus aureus biofilms using slime-producing variants and ATP-bioluminescence for automated bacterial quantification.
    Luminescence. 1999 Jan-Feb;14(1):23-31 PMID: 10398557
  37. Intravascular catheter-associated infections.
    Eur J Clin Microbiol Infect Dis. 2000 Jan;19(1):1-8 PMID: 10706172
Article Info
Journal
Antimicrobial agents and chemotherapy
Abbr.
Antimicrob Agents Chemother
ISSN
0066-4804
DOI
10.1128/AAC.45.9.2475-2479.2001
Published
2001-09-00
Pages
2475-9
Language
English
Region
United States
NLM ID
0315061
PMCID
PMC90680
Subset
IM
External Links
PubMed source DOI: 10.1128/AAC.45.9.2475-2479.2001 PMC Full Text View journal details
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