Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1997 Jan;35(1):5–10. doi: 10.1128/jcm.35.1.5-10.1997

Rapid susceptibility testing of fungi by flow cytometry using vital staining.

C Wenisch 1, K F Linnau 1, B Parschalk 1, K Zedtwitz-Liebenstein 1, A Georgopoulos 1
PMCID: PMC229504  PMID: 8968873

Abstract

A 1-h assay for antifungal susceptibility testing measuring the impairment of fungal metabolic activity was developed. Yeast viability was analyzed by flow cytometry with a novel fluorescent probe, FUN-1, which emits a red fluorescence when the yeast is metabolically active. For nine Candida albicans strains tested, this method yielded results comparable to those obtained by the standard M27 procedure for amphotericin B, flucytosine, fluconazole, and ketoconazole. Whether the flow cytometry antifungal susceptibility test results correlate with the in vivo activities of the drugs remains to determined.

Full Text

The Full Text of this article is available as a PDF (263.5 KB).

Selected References

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

  1. Anaissie E. J., Bodey G. P., Rinaldi M. G. Emerging fungal pathogens. Eur J Clin Microbiol Infect Dis. 1989 Apr;8(4):323–330. doi: 10.1007/BF01963467. [DOI] [PubMed] [Google Scholar]
  2. Barchiesi F., Colombo A. L., McGough D. A., Fothergill A. W., Rinaldi M. G. In vitro activity of itraconazole against fluconazole-susceptible and -resistant Candida albicans isolates from oral cavities of patients infected with human immunodeficiency virus. Antimicrob Agents Chemother. 1994 Jul;38(7):1530–1533. doi: 10.1128/aac.38.7.1530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Burgess S. M., Delannoy M., Jensen R. E. MMM1 encodes a mitochondrial outer membrane protein essential for establishing and maintaining the structure of yeast mitochondria. J Cell Biol. 1994 Sep;126(6):1375–1391. doi: 10.1083/jcb.126.6.1375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Calhoun D. L., Roberts G. D., Galgiani J. N., Bennett J. E., Feingold D. S., Jorgensen J., Kobayashi G. S., Shadomy S. Results of a survey of antifungal susceptibility tests in the United States and interlaboratory comparison of broth dilution testing of flucytosine and amphotericin B. J Clin Microbiol. 1986 Feb;23(2):298–301. doi: 10.1128/jcm.23.2.298-301.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Espinel-Ingroff A., Rodríguez-Tudela J. L., Martínez-Suárez J. V. Comparison of two alternative microdilution procedures with the National Committee for Clinical Laboratory Standards reference macrodilution method M27-P for in vitro testing of fluconazole-resistant and -susceptible isolates of Candida albicans. J Clin Microbiol. 1995 Dec;33(12):3154–3158. doi: 10.1128/jcm.33.12.3154-3158.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Galgiani J. N. Antifungal susceptibility tests. Antimicrob Agents Chemother. 1987 Dec;31(12):1867–1870. doi: 10.1128/aac.31.12.1867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Galgiani J. N., Reiser J., Brass C., Espinel-Ingroff A., Gordon M. A., Kerkering T. M. Comparison of relative susceptibilities of Candida species to three antifungal agents as determined by unstandardized methods. Antimicrob Agents Chemother. 1987 Sep;31(9):1343–1347. doi: 10.1128/aac.31.9.1343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ghannoum M. A., Rex J. H., Galgiani J. N. Susceptibility testing of fungi: current status of correlation of in vitro data with clinical outcome. J Clin Microbiol. 1996 Mar;34(3):489–495. doi: 10.1128/jcm.34.3.489-495.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Green L., Petersen B., Steimel L., Haeber P., Current W. Rapid determination of antifungal activity by flow cytometry. J Clin Microbiol. 1994 Apr;32(4):1088–1091. doi: 10.1128/jcm.32.4.1088-1091.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hector R. F., Braun P. C., Hart J. T., Kamarck M. E. The use of flow cytometry to monitor chitin synthesis in regenerating protoplasts of Candida albicans. J Med Vet Mycol. 1990;28(1):51–57. doi: 10.1080/02681219080000071. [DOI] [PubMed] [Google Scholar]
  11. Hitchcock C. A., Pye G. W., Troke P. F., Johnson E. M., Warnock D. W. Fluconazole resistance in Candida glabrata. Antimicrob Agents Chemother. 1993 Sep;37(9):1962–1965. doi: 10.1128/aac.37.9.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Horsburgh C. R., Jr, Kirkpatrick C. H. Long-term therapy of chronic mucocutaneous candidiasis with ketoconazole: experience with twenty-one patients. Am J Med. 1983 Jan 24;74(1B):23–29. doi: 10.1016/0002-9343(83)90511-9. [DOI] [PubMed] [Google Scholar]
  13. Mason D. J., Gant V. A. The application of flow cytometry to the estimation of bacterial antibiotic susceptibility. J Antimicrob Chemother. 1995 Aug;36(2):441–443. doi: 10.1093/jac/36.2.441. [DOI] [PubMed] [Google Scholar]
  14. Merz W. G. Candida lusitaniae: frequency of recovery, colonization, infection, and amphotericin B resistance. J Clin Microbiol. 1984 Dec;20(6):1194–1195. doi: 10.1128/jcm.20.6.1194-1195.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. O'Gorman M. R., Hopfer R. L. Amphotericin B susceptibility testing of Candida species by flow cytometry. Cytometry. 1991;12(8):743–747. doi: 10.1002/cyto.990120808. [DOI] [PubMed] [Google Scholar]
  16. Ordóez J. V., Wehman N. M. Amphotericin B susceptibility of Candida species assessed by rapid flow cytometric membrane potential assay. Cytometry. 1995 Jun 15;22(2):154–157. doi: 10.1002/cyto.990220213. [DOI] [PubMed] [Google Scholar]
  17. Pfaller M. A., Bale M., Buschelman B., Lancaster M., Espinel-Ingroff A., Rex J. H., Rinaldi M. G., Cooper C. R., McGinnis M. R. Quality control guidelines for National Committee for Clinical Laboratory Standards recommended broth macrodilution testing of amphotericin B, fluconazole, and flucytosine. J Clin Microbiol. 1995 May;33(5):1104–1107. doi: 10.1128/jcm.33.5.1104-1107.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pore R. S. Antibiotic susceptibility testing by flow cytometry. J Antimicrob Chemother. 1994 Nov;34(5):613–627. doi: 10.1093/jac/34.5.613. [DOI] [PubMed] [Google Scholar]
  19. Reinders P. P., van Vianen P. H., van der Keur M., van Engen A., Janse C. J., Tanke H. J. Computer software for testing drug susceptibility of malaria parasites. Cytometry. 1995 Mar 1;19(3):273–281. doi: 10.1002/cyto.990190312. [DOI] [PubMed] [Google Scholar]
  20. Rex J. H., Pfaller M. A., Rinaldi M. G., Polak A., Galgiani J. N. Antifungal susceptibility testing. Clin Microbiol Rev. 1993 Oct;6(4):367–381. doi: 10.1128/cmr.6.4.367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Stiller R. L., Bennett J. E., Scholer H. J., Wall M., Polak A., Stevens D. A. Susceptibility to 5-fluorocytosine and prevalence of serotype in 402 Candida albicans isolates from the United States. Antimicrob Agents Chemother. 1982 Sep;22(3):482–487. doi: 10.1128/aac.22.3.482. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wingard J. R., Merz W. G., Rinaldi M. G., Johnson T. R., Karp J. E., Saral R. Increase in Candida krusei infection among patients with bone marrow transplantation and neutropenia treated prophylactically with fluconazole. N Engl J Med. 1991 Oct 31;325(18):1274–1277. doi: 10.1056/NEJM199110313251803. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES