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. 1997 Feb;35(2):358–363. doi: 10.1128/jcm.35.2.358-363.1997

Rapid susceptibility testing of Candida albicans by flow cytometry.

S M Kirk 1, S M Callister 1, L C Lim 1, R F Schell 1
PMCID: PMC229580  PMID: 9003596

Abstract

The emerging magnitude of human fungal infections has renewed interest in developing rapid and standardized methods for susceptibility testing. We demonstrated that susceptibility testing of Candida albicans can be accomplished rapidly by using flow cytometry. Test results were available within 8 to 24 h after C. albicans isolates were incubated with amphotericin B, itraconazole, and flucytosine. This is an improvement of 24 to 60 h in the time to availability of susceptibility test results compared to the time to availability of National Committee for Clinical Laboratory Standards-recommended broth macrodilution test results. In addition, the flow cytometric endpoints, mean channel fluorescence, and number of fluorescence-labeled C. albicans cells were easy to interpret for greater sensitivity and reliability. Flow cytometry provides a more accurate means of obtaining antifungal susceptibility test results.

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Selected References

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  1. Bownds S. E., Kurzynski T. A., Norden M. A., Dufek J. L., Schell R. F. Rapid susceptibility testing for nontuberculosis mycobacteria using flow cytometry. J Clin Microbiol. 1996 Jun;34(6):1386–1390. doi: 10.1128/jcm.34.6.1386-1390.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boye E., Steen H. B., Skarstad K. Flow cytometry of bacteria: a promising tool in experimental and clinical microbiology. J Gen Microbiol. 1983 Apr;129(4):973–980. doi: 10.1099/00221287-129-4-973. [DOI] [PubMed] [Google Scholar]
  3. Diamond R. D. The growing problem of mycoses in patients infected with the human immunodeficiency virus. Rev Infect Dis. 1991 May-Jun;13(3):480–486. doi: 10.1093/clinids/13.3.480. [DOI] [PubMed] [Google Scholar]
  4. Diaper J. P., Tither K., Edwards C. Rapid assessment of bacterial viability by flow cytometry. Appl Microbiol Biotechnol. 1992 Nov;38(2):268–272. doi: 10.1007/BF00174481. [DOI] [PubMed] [Google Scholar]
  5. Dick J. D., Merz W. G., Saral R. Incidence of polyene-resistant yeasts recovered from clinical specimens. Antimicrob Agents Chemother. 1980 Jul;18(1):158–163. doi: 10.1128/aac.18.1.158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Espinel-Ingroff A., Pfaller M., Erwin M. E., Jones R. N. Interlaboratory evaluation of Etest method for testing antifungal susceptibilities of pathogenic yeasts to five antifungal agents by using Casitone agar and solidified RPMI 1640 medium with 2% glucose. J Clin Microbiol. 1996 Apr;34(4):848–852. doi: 10.1128/jcm.34.4.848-852.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Fromtling R. A., Galgiani J. N., Pfaller M. A., Espinel-Ingroff A., Bartizal K. F., Bartlett M. S., Body B. A., Frey C., Hall G., Roberts G. D. Multicenter evaluation of a broth macrodilution antifungal susceptibility test for yeasts. Antimicrob Agents Chemother. 1993 Jan;37(1):39–45. doi: 10.1128/aac.37.1.39. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gant V. A., Warnes G., Phillips I., Savidge G. F. The application of flow cytometry to the study of bacterial responses to antibiotics. J Med Microbiol. 1993 Aug;39(2):147–154. doi: 10.1099/00222615-39-2-147. [DOI] [PubMed] [Google Scholar]
  10. Georgopapadakou N. H., Tkacz J. S. The fungal cell wall as a drug target. Trends Microbiol. 1995 Mar;3(3):98–104. doi: 10.1016/s0966-842x(00)88890-3. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. Guinet R., Chanas J., Goullier A., Bonnefoy G., Ambroise-Thomas P. Fatal septicemia due to amphotericin B-resistant Candida lusitaniae. J Clin Microbiol. 1983 Aug;18(2):443–444. doi: 10.1128/jcm.18.2.443-444.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jernaes M. W., Steen H. B. Staining of Escherichia coli for flow cytometry: influx and efflux of ethidium bromide. Cytometry. 1994 Dec 1;17(4):302–309. doi: 10.1002/cyto.990170405. [DOI] [PubMed] [Google Scholar]
  15. Lim L. C., Liu Y. F., Schell K., Lovrich S. D., Callister S. M., Schell R. F. Detection of borreliacidal antibody by using acridine orange and flow cytometry. Clin Diagn Lab Immunol. 1994 Jan;1(1):44–50. doi: 10.1128/cdli.1.1.44-50.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mason D. J., Allman R., Stark J. M., Lloyd D. Rapid estimation of bacterial antibiotic susceptibility with flow cytometry. J Microsc. 1994 Oct;176(Pt 1):8–16. doi: 10.1111/j.1365-2818.1994.tb03494.x. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Mason D. J., Lopéz-Amorós R., Allman R., Stark J. M., Lloyd D. The ability of membrane potential dyes and calcafluor white to distinguish between viable and non-viable bacteria. J Appl Bacteriol. 1995 Mar;78(3):309–315. doi: 10.1111/j.1365-2672.1995.tb05031.x. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Norden M. A., Kurzynski T. A., Bownds S. E., Callister S. M., Schell R. F. Rapid susceptibility testing of Mycobacterium tuberculosis (H37Ra) by flow cytometry. J Clin Microbiol. 1995 May;33(5):1231–1237. doi: 10.1128/jcm.33.5.1231-1237.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Ordóez J. V., Wehman N. M. Rapid flow cytometric antibiotic susceptibility assay for Staphylococcus aureus. Cytometry. 1993 Oct;14(7):811–818. doi: 10.1002/cyto.990140714. [DOI] [PubMed] [Google Scholar]
  23. Polak A., Hartman P. G. Antifungal chemotherapy--are we winning? Prog Drug Res. 1991;37:181–269. doi: 10.1007/978-3-0348-7139-6_5. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. 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]
  26. Rex J. H., Rinaldi M. G., Pfaller M. A. Resistance of Candida species to fluconazole. Antimicrob Agents Chemother. 1995 Jan;39(1):1–8. doi: 10.1128/aac.39.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Steen H. B., Skarstad K., Boye E. Flow cytometry of bacteria: cell cycle kinetics and effects of antibiotics. Ann N Y Acad Sci. 1986;468:329–338. doi: 10.1111/j.1749-6632.1986.tb42050.x. [DOI] [PubMed] [Google Scholar]
  28. To W. K., Fothergill A. W., Rinaldi M. G. Comparative evaluation of macrodilution and alamar colorimetric microdilution broth methods for antifungal susceptibility testing of yeast isolates. J Clin Microbiol. 1995 Oct;33(10):2660–2664. doi: 10.1128/jcm.33.10.2660-2664.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Vanden Bossche H., Marichal P., Odds F. C. Molecular mechanisms of drug resistance in fungi. Trends Microbiol. 1994 Oct;2(10):393–400. doi: 10.1016/0966-842x(94)90618-1. [DOI] [PubMed] [Google Scholar]
  30. Vanden Bossche H., Willemsens G., Marichal P. Anti-Candida drugs--the biochemical basis for their activity. Crit Rev Microbiol. 1987;15(1):57–72. doi: 10.3109/10408418709104448. [DOI] [PubMed] [Google Scholar]
  31. van Eldere J., Joosten L., Verhaeghe V., Surmont I. Fluconazole and amphotericin B antifungal susceptibility testing by National Committee for Clinical Laboratory Standards broth macrodilution method compared with E-test and semiautomated broth microdilution test. J Clin Microbiol. 1996 Apr;34(4):842–847. doi: 10.1128/jcm.34.4.842-847.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]

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