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Susceptibility

Azole and Amphotericin B MIC Values against Aspergillus fumigatus: High Agreement between Spectrophotometric and Visual Readings Using the EUCAST EDef 9.3.2 Procedure

Julia Serrano-Lobo, Ana Gómez, Waldo Sánchez-Yebra, Miguel Fajardo, Belén Lorenzo, Ferrán Sánchez-Reus, Inmaculada Vidal, Marina Fernández-Torres, Isabel Sánchez-Romero, Carlos Ruiz de Alegría-Puig, José Luis del Pozo, Patricia Muñoz, Pilar Escribano, Jesús Guinea
Julia Serrano-Lobo
aClinical Microbiology and Infectious Diseases, Hospital General Universitario Gregorio Marañón, Madrid, Spain
bInstituto de Investigación Sanitaria Gregorio Marañón, Madrid, Spain
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Ana Gómez
aClinical Microbiology and Infectious Diseases, Hospital General Universitario Gregorio Marañón, Madrid, Spain
bInstituto de Investigación Sanitaria Gregorio Marañón, Madrid, Spain
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Waldo Sánchez-Yebra
cUnidad de Microbiología, UGC Biotecnología, Complejo Hospitalario Torrecárdenas, Almería, Spain
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Miguel Fajardo
dClinical Microbiology Department, Hospital Universitario de Badajoz, Badajoz, Spain
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Belén Lorenzo
eClinical Microbiology Department, Hospital Río Hortega, Valladolid, Spain
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Ferrán Sánchez-Reus
fClinical Microbiology, Hospital de la Santa Creu i Sant Pau, Barcelona, Spain
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  • ORCID record for Ferrán Sánchez-Reus
Inmaculada Vidal
gClinical Microbiology Department, Hospital General de Alicante, Alicante, Spain
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Marina Fernández-Torres
hClinical Microbiology Department, Hospital Universitario de Álava-BIOARABA, Vitoria, Spain
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Isabel Sánchez-Romero
iClinical Microbiology Department, Hospital Universitario Puerta de Hierro Majadahonda, Madrid, Spain
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Carlos Ruiz de Alegría-Puig
jClinical Microbiology Department, Hospital Marqués de Valdecilla-IDIVAL, Santander, Spain
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José Luis del Pozo
kClinical Microbiology and Infectious Diseases Department, Clínica Universidad de Navarra, Pamplona, Spain
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Patricia Muñoz
aClinical Microbiology and Infectious Diseases, Hospital General Universitario Gregorio Marañón, Madrid, Spain
bInstituto de Investigación Sanitaria Gregorio Marañón, Madrid, Spain
lCIBER Enfermedades Respiratorias-CIBERES (CB06/06/0058), Madrid, Spain
mMedicine Department, Faculty of Medicine, Universidad Complutense de Madrid, Madrid, Spain
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Pilar Escribano
aClinical Microbiology and Infectious Diseases, Hospital General Universitario Gregorio Marañón, Madrid, Spain
bInstituto de Investigación Sanitaria Gregorio Marañón, Madrid, Spain
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Jesús Guinea
aClinical Microbiology and Infectious Diseases, Hospital General Universitario Gregorio Marañón, Madrid, Spain
bInstituto de Investigación Sanitaria Gregorio Marañón, Madrid, Spain
lCIBER Enfermedades Respiratorias-CIBERES (CB06/06/0058), Madrid, Spain
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DOI: 10.1128/AAC.01693-20
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ABSTRACT

The EUCAST EDef 9.3.2 procedure recommends visual readings of azole and amphotericin B MICs against Aspergillus spp. Visual determination of MICs may be challenging. In this work, we aim to obtain and compare visual and spectrophotometric MIC readings of azoles and amphotericin B against Aspergillus fumigatus sensu lato isolates. A total of 847 A. fumigatus sensu lato isolates (A. fumigatus sensu stricto [n = 828] and cryptic species [n = 19]) were tested against amphotericin B, itraconazole, voriconazole, posaconazole, and isavuconazole using the EUCAST EDef 9.3.2 procedure. Isolates were classified as susceptible or resistant/non-wild type according to the 2020 updated breakpoints. The area of technical uncertainty for the azoles was defined in the updated breakpoints. Visual and spectrophotometric (fungal growth reduction of >95% compared to the control, read at 540 nm) MICs were compared. Essential (±1 2-fold dilution) and categorical agreements were calculated. Overall, high essential (97.1%) and categorical (99.6%) agreements were found. We obtained 100% categorical agreements for amphotericin B, itraconazole, and posaconazole, and consequently, no errors were found. Categorical agreements were 98.7 and 99.3% for voriconazole and isavuconazole, respectively. Most of the misclassifications for voriconazole and isavuconazole were found to be associated with MIC results falling either in the area of technical uncertainty or within one 2-fold dilution above the breakpoint. The resistance rate was slightly lower when the MICs were obtained by spectrophotometric readings. However, all relevant cyp51A mutants were correctly classified as resistant. Spectrophotometric determination of azole and amphotericin B MICs against A. fumigatus sensu lato isolates may be a convenient alternative to visual endpoint readings.

FOOTNOTES

    • Received 5 August 2020.
    • Returned for modification 27 September 2020.
    • Accepted 1 October 2020.
    • Accepted manuscript posted online 5 October 2020.
  • Supplemental material is available online only.

  • Copyright © 2020 American Society for Microbiology.

All Rights Reserved.

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Azole and Amphotericin B MIC Values against Aspergillus fumigatus: High Agreement between Spectrophotometric and Visual Readings Using the EUCAST EDef 9.3.2 Procedure
Julia Serrano-Lobo, Ana Gómez, Waldo Sánchez-Yebra, Miguel Fajardo, Belén Lorenzo, Ferrán Sánchez-Reus, Inmaculada Vidal, Marina Fernández-Torres, Isabel Sánchez-Romero, Carlos Ruiz de Alegría-Puig, José Luis del Pozo, Patricia Muñoz, Pilar Escribano, Jesús Guinea on behalf of the ASPEIN Study Group
Antimicrobial Agents and Chemotherapy Dec 2020, 65 (1) e01693-20; DOI: 10.1128/AAC.01693-20

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Azole and Amphotericin B MIC Values against Aspergillus fumigatus: High Agreement between Spectrophotometric and Visual Readings Using the EUCAST EDef 9.3.2 Procedure
Julia Serrano-Lobo, Ana Gómez, Waldo Sánchez-Yebra, Miguel Fajardo, Belén Lorenzo, Ferrán Sánchez-Reus, Inmaculada Vidal, Marina Fernández-Torres, Isabel Sánchez-Romero, Carlos Ruiz de Alegría-Puig, José Luis del Pozo, Patricia Muñoz, Pilar Escribano, Jesús Guinea on behalf of the ASPEIN Study Group
Antimicrobial Agents and Chemotherapy Dec 2020, 65 (1) e01693-20; DOI: 10.1128/AAC.01693-20
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KEYWORDS

Aspergillus fumigatus
EUCAST
azoles
amphotericin B
spectrophotometric

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