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Susceptibility

Optimal Susceptibility Testing Conditions for Detection of Azole Resistance in Aspergillus spp.: NCCLS Collaborative Evaluation

A. Espinel-Ingroff, M. Bartlett, V. Chaturvedi, M. Ghannoum, K. C. Hazen, M. A. Pfaller, M. Rinaldi, T. J. Walsh
A. Espinel-Ingroff
Medical College of Virginia of Virginia Commonwealth University, Richmond, and
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M. Bartlett
Indiana University School of Medicine, Indianapolis, Indiana;
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V. Chaturvedi
New York Health Department, Albany, New York;
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M. Ghannoum
University Hospital of Cleveland, Case Western Reserve University, Cleveland, Ohio;
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K. C. Hazen
University of Virginia Health System, Charlottesville, Virginia;
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M. A. Pfaller
University of Iowa College of Medicine, Iowa City, Iowa;
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M. Rinaldi
University of Texas Health Science Center, San Antonio, Texas; and
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T. J. Walsh
National Cancer Institute, Bethesda, Maryland
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DOI: 10.1128/AAC.45.6.1828-1835.2001
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Tables

  • Table 1.

    Selected isolates of Aspergillus spp. used in this study

    Species or study isolate no.Source no.Provider (reference no.)
    A. fumigatus
     1NCPF 7101 (AF210)a, bD. Denning (4, 5)
     2NCPF 7102 (AF294)a, bD. Denning (4)
     3NCPF 7098 (AF71)a, b, cD. Denning (4, 24)
     4NCPF 7097 (AF65)a, bD. Denning (4, 24)
     5NCPF 7100 (AF91)c, dD. Denning (4, 5, 24)
     6NCPF 7099 (AF72)dD. Denning (4, 5)
     7AZ 606eP. Verweij (NAf)
     8NIH 4215dT. Walsh (NA)
     9A 1604a, bA. Espinel-Ingroff (10, 25)
    A. flavus
     10A 1340aA. Espinel-Ingroff (7)
     11A 830aA. Espinel-Ingroff (7)
     12A 003a, bA. Espinel-Ingroff (7, 10, 25)
    A. terreus
     13A 122aA. Espinel-Ingroff (7)
     14NIH 961290aT. Walsh (NA)
     15NIH 95644aT. Walsh (NA)
    • ↵a Low itraconazole MICs (≤1.0 μg/ml) by M38-P, Etest, or other methods described in corresponding publication(s).

    • ↵b Animals and patients responded to itraconazole treatment (isolates had low itraconazole MICs [<1.0 μg/ml]).

    • ↵c These two isolates also had either a good (isolate 3) or decreased (isolate 5) response to posaconazole in a murine model.

    • ↵d Animals and patients did not respond to itraconazole treatment (isolates had high itraconazole MICs [>8 μg/ml]).

    • ↵e Voriconazole-resistant laboratory mutant.

    • ↵f NA, not available.

  • Table 2.

    Interlaboratory agreement on broth microdilution antifungal susceptibility for Aspergillusspp.a

    Antifungal agent and MIC criterion% Agreement [all isolates (itraconazole-susceptible isolates)]
    RPMIRPMI-2%M3M3-2%
    Itraconazole
     MIC-090 (92)92 (94)90 (91)85 (85)
     MIC-283 (89)88 (94)84 (92)88 (97)
    Posaconazole
     MIC-091 (91)88 (87)94 (94)86 (86)
     MIC-296 (96)95 (96)93 (99)91 (96)
    Ravuconazole
     MIC-093 (95)87 (85)94 (95)88 (89)
     MIC-288 (89)90 (89)85 (86)81 (82)
    Voriconazole
     MIC-097 (97)98 (99)99 (100)93 (92)
     MIC-288 (96)94 (95)90 (92)96 (96)
    • ↵a Species tested were A. fumigatus ( n = 9 ), A. flavus( n = 3 ), and A. terreus ( n = 3 ); 48-h MICs are shown.

  • Table 3.

    Itraconazole MICs for Aspergillus spp. in eight laboratoriesa

    Isolate(s) (no. tested) and MIC criterionMIC range (MIC90 or modal MIC)b
    RPMIRPMI-2%M3M3-2%
    Itraconazole susceptible (12)
     MIC-00.03–2 (1)0.03–4 (0.5)0.06–>8 (1)0.06–>8 (2)
     MIC-20.01–1 (0.2)<0.008–0.5 (0.12)<0.008–1 (0.2)0.01–1 (0.2)
    Itraconazole resistant (isolate no. 5)
     MIC-08–>8 (>8)8–>8 (>8)2–>8 (>8)2–>8 (>8)
     MIC-20.06–>8 (2)0.06–>8 (1)<0.008–>8 (1)<0.008–>8 (1)
    Itraconazole resistant (isolate no. 6)
     MIC-01–>8 (>8)1–>8 (>8)2–>8 (>8)1–>8 (>8)
     MIC-20.12–>8 (1)0.12–>8 (0.5)1–>8 (1)0.2–>8 (1)
    Voriconazole-resistant mutant (1)
     MIC-00.5–>8 (2)0.5–>8 (0.5)1–>8 (>8)1–>8 (>8)
     MIC-20.06–0.5 (0.2)0.03–0.5 (0.12)0.06–0.5 (0.2)0.01–0.2 (0.06)
    • ↵a Species tested were A. fumigatus ( n = 9 ), A. flavus( n = 3 ), and A. terreus ( n = 3 ); see Table 1 for reported data regarding isolates 5 and 6; 48-h MICs are shown.

    • ↵b MICs are reported in micrograms per milliliter; the MIC90 is reported for susceptible isolates and the modal MIC is reported for single resistant isolates.

  • Table 4.

    Posaconazole (SCH56592) MICs for Aspergillusspp. in eight laboratoriesa

    Isolate(s) (no. tested) and MIC criterionMIC range (MIC90 or modal MIC)b
    RPMIRPMI-2%M3M3-2%
    Itraconazole susceptible (12)
     MIC-00.01–0.5 (0.2)0.03–2 (0.2)0.03–0.5 (0.2)0.03–>8 (0.5)
     MIC-20.03–0.12 (0.06)0.008–0.12 (0.06)0.008–0.12 (0.06)0.03–0.06 (0.06)
    Itraconazole resistant (isolate no. 5)
     MIC-00.2–1 (1)0.2–2 (0.5)0.5–2 (0.5)0.2–2 (0.5)
     MIC-20.03–0.5 (0.2)0.03–0.2 (0.12)<0.008–0.5 (0.5)<0.008–0.5 (0.2)
    Itraconazole resistant (isolate no. 6)
     MIC-00.2–1 (0.5)0.2–1 (0.5)0.12–2 (1)0.12–>8 (0.5)
     MIC-20.06–0.5 (0.12)0.01–0.2 (0.12)0.01–1 (0.2)0.03–0.5 (0.2)
    Voriconazole-resistant mutant
     MIC-00.12–>8 (0.2)0.06–2 (0.12)0.5–8 (0.5)0.12–2 (0.2)
     MIC-20.03–0.12 (0.06)0.03–1 (0.03)0.03–0.12 (0.03)0.03–0.06 (0.03)
    • ↵a Species tested were A. fumigatus ( n = 9 ), A. flavus( n = 3 ), and A. terreus ( n = 3 ); see Table 1 for reported data regarding isolates 5 and 6; 48-h MICs are shown.

    • ↵b MICs are reported in micrograms per milliliter; the MIC90 is reported for susceptible isolates and the model MIC is reported for single resistant isolates.

  • Table 5.

    Ravuconazole (BMS-207147) MICs for Aspergillusspp. in eight laboratoriesa

    Isolate(s) (no. tested) and MIC criterionMIC range (MIC90 or modal MIC)b
    RPMIRPMI-2%M3M3-2%
    Itraconazole susceptible (12)
     MIC-00.12–4 (2)0.12–>8 (1)0.2–>8 (4)0.2–>8 (4)
     MIC-20.01–1 (1)0.01–0.5 (0.5)0.01–4 (1)0.06–1 (1)
    Itraconazole resistant (isolate no. 5)
     MIC-00.2–0.5 (0.2)0.12–1 (0.2)0.2–8 (0.5)0.12–4 (1)
     MIC-20.01–0.2 (0.12)0.03–0.12 (0.06)<0.008–0.5 (0.12)0.008–0.5 (0.06)
    Itraconazole resistant (isolate no. 6)
     MIC-02–8 (2)1–>8 (2)1–>8 (8)2–>8 (8)
     MIC-20.2–4 (0.5)0.12–1 (1)0.2–4 (2)0.062–8 (2)
    Voriconazole-resistant mutant (1)
     MIC-02–>8 (4)1–4 (2)4–>8 (>8)2–>8 (8)
     MIC-20.12–4 (1)0.06–1 (0.5)0.06–2 (0.5)0.03–1 (0.2)
    • ↵a Species tested were A. fumigatus ( n = 9 ), A. flavus( n = 3 ), and A. terreus ( n = 3 ); see Table 1 for reported data regarding isolates 5 and 6; 48-h MICs are shown.

    • ↵b MICs are reported in micrograms per milliliter; the MIC90 is reported for susceptible isolates and the modal MIC is reported for single resistant isolates.

  • Table 6.

    Voriconazole MICs for Aspergillus spp. in eight laboratoriesa

    Isolate(s) (no. tested) and MIC criterionMIC range (MIC90 or modal MIC)b
    RPMIRPMI-2%M3M3-2%
    Itraconazole susceptible (12)
     MIC-00.12–2 (1)0.2–2 (1)0.5–8 (4)0.2–>8 (4)
     MIC-20.01–2 (0.5)<0.008–1 (1)<0.008–1 (1)0.12–1 (0.5)
    Itraconazole resistant (isolate no. 5)
     MIC-00.2–0.5 (0.2)0.2–0.5 (0.2)0.2–2 (0.5)0.2–1 (0.5)
     MIC-2 (0.12)0.03–0.2 (0.12)0.03–0.2 (0.06)<0.008–0.2 (0.12)0.03–0.12 (0.12)
    Itraconazole resistant (isolate no. 6)
     MIC-01–2 (1)0.5–2 (1)2–4 (4)2–8 (2)
     MIC-20.12–0.5 (0.5)0.12–1 (0.2)0.5–2 (1)0.2–1 (0.5)
    Voriconazole-resistant mutant (1)
     MIC-02–>8 (4)2–8 (4)4–>8 (8)4–>8 (8)
     MIC-20.5–4 (0.5)0.2–1 (1)0.2–2 (0.5)0.2–1 (0.5)
    • ↵a Species tested were A. fumigatus ( n = 9 ), A. flavus( n = 3 ), and A. terreus ( n = 3 ); 48-h MICs are shown.

    • ↵b MICs are reported in micrograms per milliliter; the MIC90 is reported for susceptible isolates and the modal MIC is reported for single resistant isolates.

  • Table 7.

    Triazole MIC ranges for the reference A. flavus ATCC 204304 isolate in eight laboratories

    Antifungal agent and MIC criterionMIC rangea
    RPMIRPMI-2%M3M3-2%
    Itraconazole
     MIC-00.2–0.50.12–0.50.5–10.5–1
     MIC-2b0.06–0.20.06–0.50.06–0.20.06–0.5
    Posaconazole
     MIC-00.06–0.50.06–0.50.12–0.20.12
     MIC-20.03–0.120.03–0.060.01–0.120.03–0.06
    Ravuconazole
     MIC-00.5–40.5–21–40.5–4
     MIC-20.12–10.06–10.12–10.06–2
    Voriconazole
     MIC-00.5–40.5–20.5–>80.5–4
     MIC-20.12–0.50.06–0.50.12–10.12–2
    • ↵a Forty-eight-hour MICs are shown; MICs are reported in micrograms per milliliter.

    • ↵b The reference MIC-2 range is 0.12 to 0.5 μg/ml (10).

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Optimal Susceptibility Testing Conditions for Detection of Azole Resistance in Aspergillus spp.: NCCLS Collaborative Evaluation
A. Espinel-Ingroff, M. Bartlett, V. Chaturvedi, M. Ghannoum, K. C. Hazen, M. A. Pfaller, M. Rinaldi, T. J. Walsh
Antimicrobial Agents and Chemotherapy Jun 2001, 45 (6) 1828-1835; DOI: 10.1128/AAC.45.6.1828-1835.2001

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Optimal Susceptibility Testing Conditions for Detection of Azole Resistance in Aspergillus spp.: NCCLS Collaborative Evaluation
A. Espinel-Ingroff, M. Bartlett, V. Chaturvedi, M. Ghannoum, K. C. Hazen, M. A. Pfaller, M. Rinaldi, T. J. Walsh
Antimicrobial Agents and Chemotherapy Jun 2001, 45 (6) 1828-1835; DOI: 10.1128/AAC.45.6.1828-1835.2001
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KEYWORDS

antifungal agents
Aspergillus
triazoles

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