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Chapter 8.10 : A S T

Editor: Amy L. Leber1
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Affiliations: 1: Editor in chief, Department of Laboratory Medicine, Nationwide Children’s Hospital, Columbus, Ohio
Content Type: Reference
Publication Year: 2016

Category: Clinical Microbiology

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Abstract:

Until recently, antifungal susceptibility testing (AFST) has lagged behind its antibacterial counterpart. A number of important achievements helped propel the antifungal susceptibility field. These achievements include publication of the approved reference method for broth dilution AFST of yeast, CLSI document M27-A3 ( ); publication of interpretive breakpoints using this method ( ); the recommended use of AFST during selection of therapy of patients with candidemia and hematogenously disseminated candidiasis as outlined by the published guidelines for the treatment of candidiasis ( ); publication of a reference method for broth dilution AFST of conidium-forming filamentous fungi, CLSI document M38-A2 ( ); and recent development and publication of AFST methods for dermatophytes ( ). The procedures described below are based on CLSI documents M27-A3, M27-S4, and M38-A2.

Citation: Leber A. 2016. A S T, p 8.10.1.1-8.10.4.1. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch8.10
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References

/content/book/10.1128/9781555818814.chap8.10
1. CLSI. 2008. Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts, 3rd ed. Approved standard M27-A3. CLSI, Wayne, PA.
2. CLSI. 2012. Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts, 4th ed. Informational supplement M27-S4. CLSI, Wayne, PA.
3. Pappas PG, Rex JR, Sobel JD, Filler SG, Dismukes WE, Walsh TJ, Edwards JE. 2004. Guidelines for treatment of candidiasis. Clin Infect Dis 38: 161– 189.
4. CLSI. 2008. Reference Method for Broth Dilution Antifungal Susceptibility Testing of Filamentous Fungi. Approved standard M38-A2. CLSI, Wayne, PA.
5. Jessup CJ, Warner J, Isham N, Hasan I, Ghannoum MA. 2000. Antifungal susceptibility testing of dermatophytes: establishing a medium for inducing conidial growth and evaluation of susceptibility of clinical isolates. J Clin Microbiol 38: 341– 344.
6. Pfaller MA, Boyken L, Hollis RJ, Kroeger J, Messer SA, Tendolkar S, Diekema DJ. 2008. In vitro susceptibility of invasive isolates of Candida spp. to anidulafungin, caspofungin, and micafungin: six years of global surveillance. J Clin Microbiol 46: 150– 156.
7. Ghannoum MA, Ibrahim AS, Fu Y, Shafiq MC, Edwards JE, Criddle RS. 1992. Susceptibility testing of Cryptococcus neoformans: a microdilution technique. J Clin Microbiol 30: 2881– 2886.
8. Pfaller MA, Boyken LB, Hollis RJ, Kroeger J, Messer SA, Tendolkar S, Diekema DJ.. 2011. Validation of 24-hour posaconazole and voriconazole MIC readings versus the CLSI 48-hour broth microdilution reference method: application of epidemiological cutoff values to results from a global Candida antifungal surveillance program. J Clin Microbiol. 49: 1274– 1279.
9. Pfaller MA, Andes D, Arendrup MC, Diekema DJ, Espinel-Ingroff A, Alexander BD, Brown SD, Chaturvedi V, Fowler CL, Ghannoum MA, Johnson EM, Knapp CC, Motyl MR, Ostrosky-Zeichner L, Walsh TJ.. 2011. Clinical breakpoints for voriconazole and Candida spp. revisited: review of microbiologic, molecular, pharmacodynamic, and clinical data as they pertain to the development of species-specific interpretive criteria. Diagn Microbiol Infect Dis 70: 330– 343.
10. Pfaller MA, Andes D, Diekema DJ, Espinel-Ingroff A, Sheehan D; ClSI Subcommittee for Antifungal Susceptibility Testing. 2010. Wild-type MIC distributions, epidemiological cutoff values and species-specific clinical breakpoints for fluconazole and Candida: time for harmonization of CLSI and EUCAST broth microdilution methods. Drug Resist Updat 13: 180– 195.
11. Pfaller MA, Diekema DJ, Andes D, Arendrup MC, Brown SD, Lockhart SR, Motyl M, Perlin DS; CLSI Subcommittee for Antifungal Testing. 2011. Clinical breakpoints for the echinocandins and Candida revisited: integration of molecular, clinical, and Microbiological data to arrive at species-specific interpretive criteria. Drug Resist Updat 14: 164– 176.
12. Arendrup MC, Perlin DS, Jensen RH, Howard SJ, Goodwin J, Hope W. 2012. Differential in vivo activities of anidulafungin, caspofungin and micafungin against Candida glabrata with and without FKS resistance mutations. Antimicrob Agents Chemother 56: 2435– 2442.
13. Pfaller MA, Diekema DJ, Ostrosky-Zeichner L, Rex JH, Alexander BD, Andes D, Brown SD, Chaturvedi V, Ghannoum MA, Knapp CC, Sheehan DJ, Walsh TJ. 2008. Correlation of MIC with outcome for Candida species tested against caspofungin, anidulafungin, and micafungin: analysis and proposal for interpretive MIC breakpoints. J Clin Microbiol 46: 2620– 2629.
14. Pfaller MA, Diekema DJ, Rex JH, Espinel-Ingroff A, Johnson EM, Andes D, Chaturvedi V, Ghannoum MA, Odds FC, Rinaldi MG, Sheehan DJ, Troke P, Walsh TJ, Warnock DW. 2006. Correlation of MIC with outcome for Candida species tested against voriconazole: analysis and proposal for interpretive breakpoints. J Clin Microbiol 44: 819– 826.
15. Rex JH, Pfaller MA, Galgiani JN, Bartlett MS, Espinel-Ingroff A, Ghannoum MA, Lancaster M, Odds FC, Rinaldi MG, Walsh TJ, Barry AL. 1997. 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 24: 235– 247.
1. CLSI. 2009. Method for Antifungal Disk Diffusion Susceptibility Testing of Yeasts: Approved Guideline, 2nd ed. Approved standard M44-A2. CLSI, Wayne, PA.
2. CLSI. 2009. Zone Diameter Interpretive Standards, Corresponding Minimal Inhibitory Concentration (MIC) Interpretive Breakpoints, and Quality Control Limits for Antifungal Disk Diffusion Susceptibility Testing of Yeasts; Third Information Supplement. Informational supplement M44-S3. CLSI, Wayne, PA.
3. CLSI. 2010. Method for Antifungal Disk Diffusion Susceptibility Testing of Nondermatophyte Filamentous Fungi. Approved guideline 51-A. CLSI, Wayne, PA,
4. Pfaller MA, Diekema DJ, Ostrosky-Zeichner L, Rex JH, Alexander BD, Andes D, Brown SD, Chaturvedi V, Ghannoum MA, Knapp CC, Sheehan DJ, Walsh TJ. 2008. Correlation of MIC with outcome for Candida species tested against caspofungin, anidulafungin, and micafungin: analysis and proposal for interpretive MIC breakpoints. J Clin Microbiol 46: 2620– 2629.
5. Brown SD, Traczewski MM. 2008. Caspofungin disk diffusion breakpoints and quality control. J Clin Microbiol 46: 1927– 1929.
6. Rex JH, Pfaller MA, Galgiani JN, Bartlett MS, Espinel-Ingroff A, Ghannoum MA, Lancaster M, Odds FC, Rinaldi MG, Walsh TJ, Barry AL. 1997. 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 24: 235– 247.
7. Pfaller MA, Diekema DJ, Sheehan DJ. 2006. Interpretive breakpoints for fluconazole and Candida revisited: a blueprint for the future of antifungal susceptibility testing. Clin Microbiol Rev 19: 435– 447.
8. Barry AL, Pfaller MA, Rennie RP, Fuchs PC, Brown SD. 2002. Precision and accuracy of fluconazole susceptibility testing by broth microdilution, Etest, and disk diffusion methods. Antimicrob Agents Chemother 46: 1781– 1784.
9. Pfaller MA, Hazen KC, Messer SA, Boyken L, Tendolkar S, Hollis RJ, Diekema DJ. 2004. Comparison of results of fluconazole disk diffusion testing for Candida species with results from a central reference laboratory in the ARTEMIS global antifungal surveillance program. J Clin Microbiol 42: 3607– 3612.
10. Pfaller MA, Diekema DJ, Rex JH, Espinel-Ingroff A, Johnson EM, Andes D, Chaturvedi V, Ghannoum MA, Odds FC, Rinaldi MG, Sheehan DJ, Troke P, Walsh TJ, Warnock DW. 2006. Correlation of MIC with outcome for Candida species tested against voriconazole: analysis and proposal for interpretive breakpoints. J Clin Microbiol 44: 819– 826.
11. Barry A, Bille J, Brown S, Ellis D, Meis J, Pfaller M, Rennie R, Rinaldi M, Rogers T, Traczewski M. 2003. Quality control limits for fluconazole disk susceptibility tests on Mueller-Hinton agar with glucose and methylene blue. J Clin Microbiol 41: 3410– 3412.
12. Brown S, Traczewski M. 2007. Quality control limits for posaconazole disk susceptibility tests on Mueller-Hinton agar with glucose and methylene blue. J Clin Microbiol 45: 222– 223.
13. Pfaller MA, Barry A, Bille J, Brown S, Ellis D, Meis JF, Rennie R, Rinaldi M, Rogers T, Traczewski M. 2004. Quality control limits for voriconazole disk susceptibility tests on Mueller-Hinton agar with glucose and methylene blue. J Clin Microbiol 42: 1716– 1718.
14. Espinel-Ingroff A, Arthington-Skaggs B, Iqbal N, Ellis D, Pfaller MA, Messer S, Rinaldi M, Fothergill A, Gibbs DL, Wang A. 2007. Multicenter evaluation of a new disk agar diffusion method for susceptibility testing of filamentous fungi with voriconazole, posaconazole, itraconazole, amphotericin B, and caspofungin. J Clin Microbiol 45: 1811– 1820.
1. Colombo AL, Barchiesi F, McGough DA, Fothergill AW, Rinaldi MG. 1995. Evaluation of the E test system versus a microtitre broth method for antifungal susceptibility testing of yeasts against fluconazole and itraconazole. J Antimicrob Chemother 36: 93– 100.
1. Kahlmeter G, Brown DF, Goldstein FW, MacGowan AP, Mouton JW, Osterlund A, Rodloff A, Steinbakk M, Urbaskova P, Vatopoulos A. 2003. European harmonization of MIC breakpoints for antimicrobial susceptibility testing of bacteria. J Antimicrob Chemother 52: 145– 148.
2. Pfaller MA, Diekema DJ, Ghannoum MA, Rex JH, Alexander BD, Andes D, Brown SD, Chaturvedi V, Espinel-Ingroff A, Fowler CL, Johnson EM, Knapp CC, Motyl MR, Ostrosky-Zeichner L, Sheehan DJ, and Walsh TJ for the Clinical and Laboratory Standards Institute Antifungal Testing Subcommittee. 2009. Wild-Type MIC distribution and epidemiological cutoff values for Aspergillus fumigatus and three triazoles as determined by the Clinical and Laboratory Standards Institute Broth Microdilution Methods. J Clin Microbiol 47: 3142– 3146.
3. Pfaller MA, Andes D, Diekema Espinel-Ingroff A, Sheehan D The CLSI Subcommittee for Antifungal Susceptibility Testing. 2010. Wild-type MIC distributions, epidemiological cutoff values and species-specific clinical breakpoints for fluconazole and candida: time for harmonization of CLSI and EUCAST broth microdilution methods. Drug Resis Updates 13: 180– 195.

Tables

Antifungal Susceptibility Testing
Citation: Leber A. 2016. A S T, p 8.10.1.1-8.10.4.1. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch8.10
/content/10.1128/9781555818814.chap8.10.table8.10.1-1
Table 8.10.1–1
Dilution procedure for water-soluble drugs
Generic image for table
Table 8.10.1–1

Dilution procedure for water-soluble drugs

Citation: Leber A. 2016. A S T, p 8.10.1.1-8.10.4.1. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch8.10
/content/10.1128/9781555818814.chap8.10.table8.10.1-2
Table 8.10.1–2
Dilution procedure for water-insoluble drugs
Generic image for table
Table 8.10.1–2

Dilution procedure for water-insoluble drugs

Citation: Leber A. 2016. A S T, p 8.10.1.1-8.10.4.1. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch8.10
/content/10.1128/9781555818814.chap8.10.table8.10.1-3
Table 8.10.1–3
Recommended 24- and 48-h MIC limits for two QC strains for broth microdilution ( )
Generic image for table
Table 8.10.1–3

Recommended 24- and 48-h MIC limits for two QC strains for broth microdilution ( )

Citation: Leber A. 2016. A S T, p 8.10.1.1-8.10.4.1. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch8.10
/content/10.1128/9781555818814.chap8.10.table8.10.1-4
Table 8.10.1–4
Dilution procedure for water-insoluble drugs used in broth microdilution susceptibility tests for nondermatophyte filamentous moulds
Generic image for table
Table 8.10.1–4

Dilution procedure for water-insoluble drugs used in broth microdilution susceptibility tests for nondermatophyte filamentous moulds

Citation: Leber A. 2016. A S T, p 8.10.1.1-8.10.4.1. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch8.10
/content/10.1128/9781555818814.chap8.10.table8.10.1-5
Table 8.10.1–5
Dilution procedure for water-insoluble drugs used in broth dilution susceptibility tests for dermatophyte isolates
Generic image for table
Table 8.10.1–5

Dilution procedure for water-insoluble drugs used in broth dilution susceptibility tests for dermatophyte isolates

Citation: Leber A. 2016. A S T, p 8.10.1.1-8.10.4.1. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch8.10
/content/10.1128/9781555818814.chap8.10.table8.10.1-6
Table 8.10.1–6
Dilution procedure for water-soluble drugs used in broth dilution susceptibility tests of dermatophyte and nondermatophyte filamentous moulds
Generic image for table
Table 8.10.1–6

Dilution procedure for water-soluble drugs used in broth dilution susceptibility tests of dermatophyte and nondermatophyte filamentous moulds

Citation: Leber A. 2016. A S T, p 8.10.1.1-8.10.4.1. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch8.10
/content/10.1128/9781555818814.chap8.10.table8.10.1-7
Table 8.10.1–7
Interpretive guidelines for susceptibility testing of spp. and selected azoles after 24-h incubation
Generic image for table
Table 8.10.1–7

Interpretive guidelines for susceptibility testing of spp. and selected azoles after 24-h incubation

Citation: Leber A. 2016. A S T, p 8.10.1.1-8.10.4.1. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch8.10
/content/10.1128/9781555818814.chap8.10.table8.10.1-8
Table 8.10.1–8
Interpretive guidelines for susceptibility testing of spp. and echinocandins ( )
Generic image for table
Table 8.10.1–8

Interpretive guidelines for susceptibility testing of spp. and echinocandins ( )

Citation: Leber A. 2016. A S T, p 8.10.1.1-8.10.4.1. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch8.10
/content/10.1128/9781555818814.chap8.10.table8.10.2-1
Table 8.10.2–1
Recommended quality control zone diameter (mm) ranges for testing yeast isolates
Generic image for table
Table 8.10.2–1

Recommended quality control zone diameter (mm) ranges for testing yeast isolates

Citation: Leber A. 2016. A S T, p 8.10.1.1-8.10.4.1. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch8.10
/content/10.1128/9781555818814.chap8.10.table8.10.2-2
Table 8.10.2–2
Recommended QC zone diameter (mm) ranges for testing filamentous fungi
Generic image for table
Table 8.10.2–2

Recommended QC zone diameter (mm) ranges for testing filamentous fungi

Citation: Leber A. 2016. A S T, p 8.10.1.1-8.10.4.1. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch8.10
/content/10.1128/9781555818814.chap8.10.table8.10.2-3
Table 8.10.2–3
Zone diameter interpretive standards and corresponding MIC breakpoints for select antifungal agents against spp.
Generic image for table
Table 8.10.2–3

Zone diameter interpretive standards and corresponding MIC breakpoints for select antifungal agents against spp.

Citation: Leber A. 2016. A S T, p 8.10.1.1-8.10.4.1. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch8.10

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