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. 1992 Dec;30(12):3138–3145. doi: 10.1128/jcm.30.12.3138-3145.1992

Collaborative comparison of broth macrodilution and microdilution antifungal susceptibility tests.

A Espinel-Ingroff 1, C W Kish Jr 1, T M Kerkering 1, R A Fromtling 1, K Bartizal 1, J N Galgiani 1, K Villareal 1, M A Pfaller 1, T Gerarden 1, M G Rinaldi 1, et al.
PMCID: PMC270602  PMID: 1452697

Abstract

A collaborative comparison of macro- and microdilution antifungal susceptibility tests was performed in five laboratories. MICs of amphotericin B, fluconazole, flucytosine, and ketoconazole were determined in all five centers against 95 coded isolates of Candida spp., Cryptococcus neoformans, and Torulopsis glabrata. A standard protocol with the following National Committee for Clinical Laboratory Standards Subcommittee on Antifungal Susceptibility Testing recommendations was used: an inoculum standardized by spectrophotometer, buffered (RPMI 1640) medium (pH 7.0), incubation at 35 degrees C, and an additive drug dilution procedure. Two inoculum sizes were tested (1 x 10(4) to 5 x 10(3) to 2.5 x 10(3) CFU/ml) and three scoring criteria were evaluated for MIC endpoint determinations, which were scored as 0 (optically clear), < or = 1 (slightly hazy turbidity), and < or = 2 (prominent decrease in turbidity compared with that of the growth control). Overall intra- and interlaboratory reproducibility was optimal with the low-density inoculum, the second-day readings, and MICs scored as either 1 or 2. The microdilution MICs demonstrated interlaboratory agreement with most of the four drugs higher than or similar to that of the macrodilution MICs. In general, there was good interlaboratory agreement with amphotericin B, fluconazole, and flucytosine; ketoconazole gave more variable results.

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Selected References

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  1. Bruatto M., Vidotto V., Marinuzzi G., Raiteri R., Sinicco A. Candida albicans biotypes in human immunodeficiency virus type 1-infected patients with oral candidiasis before and after antifungal therapy. J Clin Microbiol. 1991 Apr;29(4):726–730. doi: 10.1128/jcm.29.4.726-730.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Coleman T., Madassery J. V., Kobayashi G. S., Nahm M. H., Little J. R. New fluorescence assay for the quantitation of fungi. J Clin Microbiol. 1989 Sep;27(9):2003–2007. doi: 10.1128/jcm.27.9.2003-2007.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Doern G. V., Tubert T. A., Chapin K., Rinaldi M. G. Effect of medium composition on results of macrobroth dilution antifungal susceptibility testing of yeasts. J Clin Microbiol. 1986 Oct;24(4):507–511. doi: 10.1128/jcm.24.4.507-511.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Eng C., Valenstein P. Standardization of yeast inocula with an electronic impedance counter. J Clin Microbiol. 1989 Oct;27(10):2397–2399. doi: 10.1128/jcm.27.10.2397-2399.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Espinel-Ingroff A., Kerkering T. M., Goldson P. R., Shadomy S. Comparison study of broth macrodilution and microdilution antifungal susceptibility tests. J Clin Microbiol. 1991 Jun;29(6):1089–1094. doi: 10.1128/jcm.29.6.1089-1094.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fan-Havard P., Capano D., Smith S. M., Mangia A., Eng R. H. Development of resistance in candida isolates from patients receiving prolonged antifungal therapy. Antimicrob Agents Chemother. 1991 Nov;35(11):2302–2305. doi: 10.1128/aac.35.11.2302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Galgiani J. N. Susceptibility of Candida albicans and other yeasts to fluconazole: relation between in vitro and in vivo studies. Rev Infect Dis. 1990 Mar-Apr;12 (Suppl 3):S272–S275. doi: 10.1093/clinids/12.supplement_3.s272. [DOI] [PubMed] [Google Scholar]
  10. Hector R. F., Zimmer B. L., Pappagianis D. Microtiter method for MIC testing with spherule-endospore-phase Coccidioides immitis. J Clin Microbiol. 1988 Dec;26(12):2667–2668. doi: 10.1128/jcm.26.12.2667-2668.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Holmberg K., Meyer R. D. Fungal infections in patients with AIDS and AIDS-related complex. Scand J Infect Dis. 1986;18(3):179–192. doi: 10.3109/00365548609032326. [DOI] [PubMed] [Google Scholar]
  12. Johnson E. M., Richardson M. D., Warnock D. W. In-vitro resistance to imidazole antifungals in Candida albicans. J Antimicrob Chemother. 1984 Jun;13(6):547–558. doi: 10.1093/jac/13.6.547. [DOI] [PubMed] [Google Scholar]
  13. Klein R. S., Harris C. A., Small C. B., Moll B., Lesser M., Friedland G. H. Oral candidiasis in high-risk patients as the initial manifestation of the acquired immunodeficiency syndrome. N Engl J Med. 1984 Aug 9;311(6):354–358. doi: 10.1056/NEJM198408093110602. [DOI] [PubMed] [Google Scholar]
  14. Kobayashi G. S., Spitzer E. D. Testing of organisms for susceptibility to triazoles: is it justified? Eur J Clin Microbiol Infect Dis. 1989 May;8(5):387–389. doi: 10.1007/BF01964051. [DOI] [PubMed] [Google Scholar]
  15. Korting H. C., Ollert M., Georgii A., Fröschl M. In vitro susceptibilities and biotypes of Candida albicans isolates from the oral cavities of patients infected with human immunodeficiency virus. J Clin Microbiol. 1988 Dec;26(12):2626–2631. doi: 10.1128/jcm.26.12.2626-2631.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. McIntyre K. A., Galgiani J. N. In vitro susceptibilities of yeasts to a new antifungal triazole, SCH 39304: effects of test conditions and relation to in vivo efficacy. Antimicrob Agents Chemother. 1989 Jul;33(7):1095–1100. doi: 10.1128/aac.33.7.1095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Merz W. G., Khazan U., Jabra-Rizk M. A., Wu L. C., Osterhout G. J., Lehmann P. F. Strain delineation and epidemiology of Candida (Clavispora) lusitaniae. J Clin Microbiol. 1992 Feb;30(2):449–454. doi: 10.1128/jcm.30.2.449-454.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Odds F. C. Quantitative microculture system with standardized inocula for strain typing, susceptibility testing, and other physiologic measurements with Candida albicans and other yeasts. J Clin Microbiol. 1991 Dec;29(12):2735–2740. doi: 10.1128/jcm.29.12.2735-2740.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pfaller M. A., Burmeister L., Bartlett M. S., Rinaldi M. G. Multicenter evaluation of four methods of yeast inoculum preparation. J Clin Microbiol. 1988 Aug;26(8):1437–1441. doi: 10.1128/jcm.26.8.1437-1441.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Pfaller M. A., Rinaldi M. G., Galgiani J. N., Bartlett M. S., Body B. A., Espinel-Ingroff A., Fromtling R. A., Hall G. S., Hughes C. E., Odds F. C. Collaborative investigation of variables in susceptibility testing of yeasts. Antimicrob Agents Chemother. 1990 Sep;34(9):1648–1654. doi: 10.1128/aac.34.9.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Radetsky M., Wheeler R. C., Roe M. H., Todd J. K. Microtiter broth dilution method for yeast susceptibility testing with validation by clinical outcome. J Clin Microbiol. 1986 Oct;24(4):600–606. doi: 10.1128/jcm.24.4.600-606.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Smith K. J., Warnock D. W., Kennedy C. T., Johnson E. M., Hopwood V., Van Cutsem J., Vanden Bossche H. Azole resistance in Candida albicans. J Med Vet Mycol. 1986 Apr;24(2):133–144. [PubMed] [Google Scholar]
  23. Soll D. R., Galask R., Isley S., Rao T. V., Stone D., Hicks J., Schmid J., Mac K., Hanna C. Switching of Candida albicans during successive episodes of recurrent vaginitis. J Clin Microbiol. 1989 Apr;27(4):681–690. doi: 10.1128/jcm.27.4.681-690.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Warnock D. W., Burke J., Cope N. J., Johnson E. M., von Fraunhofer N. A., Williams E. W. Fluconazole resistance in Candida glabrata. Lancet. 1988 Dec 3;2(8623):1310–1310. doi: 10.1016/s0140-6736(88)92919-4. [DOI] [PubMed] [Google Scholar]
  25. Warnock D. W. Itraconazole and fluconazole: new drugs for deep fungal infection. J Antimicrob Chemother. 1989 Sep;24(3):275–277. doi: 10.1093/jac/24.3.275. [DOI] [PubMed] [Google Scholar]
  26. Wey S. B., Mori M., Pfaller M. A., Woolson R. F., Wenzel R. P. Hospital-acquired candidemia. The attributable mortality and excess length of stay. Arch Intern Med. 1988 Dec;148(12):2642–2645. doi: 10.1001/archinte.148.12.2642. [DOI] [PubMed] [Google Scholar]

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