EUCAST Definitive Document EDef 7.1: method for the determination of broth dilution MICs of antifungal agents for fermentative yeasts

Rodríguez-Tudela, Juan Luis; Arendrup, Maiken Cavling; Barchiesi, Francesco; Billé, Jacques; Chryssanthou, Erja; Cuenca-Estrella, M.; Dannaoui, Éric; Denning, David W.; Donnelly, J. Peter; Dromer, Françoise; Fegeler, W.; Lass‐Flörl, Cornelia; Moore, Caroline B.; Richardson, Malcolm; Sandven, Per; Velegraki, Aristea; Verweij, Paul E.

doi:10.1111/j.1469-0691.2007.01935.x

Cited by 464 publications

(248 citation statements)

References 14 publications

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“…Susceptibility testing was performed according to the protocol in EUCAST definitive document EDef 7.1 (45). Manufacturers and stock concentrations of reagents were as follows: dimethyl sulfoxide (DMSO) (D8779), Sigma-Aldrich; fluconazole, Pfizer A/S, Ballerup, Denmark (10,000 g/ml); amphotericin B (A2411), Sigma-Aldrich, Vallensbaek Strand, Denmark (5,000 g/ml in DMSO); caspofungin, Merck, Sharp and Dohme, Glostrup, Denmark (5,000 g/ml in DMSO); itraconazole, Janssen-Cilag, Birkerød, Denmark (5,000 g/ml in DMSO); and voriconazole, Pfizer A/S, Ballerup, Denmark (5,000 g/ml in DMSO).…”

Section: Methodsmentioning

confidence: 99%

Diagnostic Issues, Clinical Characteristics, and Outcomes for Patients with Fungemia

et al. 2011

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This study investigated microbiological, clinical, and management issues and outcomes for Danish fungemia patients. Isolates and clinical information were collected at six centers. A total of 334 isolates, 316 episodes, and 305 patients were included, corresponding to 2/3 of the national episodes. Blood culture positivity varied by system, species, and procedure. Thus, cases with concomitant bacteremia were reported less commonly by BacT/Alert than by the Bactec system (9% [11/124 cases] versus 28% [53/192 cases]; P < 0.0001), and cultures with Candida glabrata or those drawn via arterial lines needed longer incubation. Species distribution varied by age, prior antifungal treatment (57% occurrence of C. glabrata, Saccharomyces cerevisiae, or C. krusei in patients with prior antifungal treatment versus 28% occurrence in those without it; P ‫؍‬ 0.007), and clinical specialty (61% occurrence of C. glabrata or C. krusei in hematology wards versus 27% occurrence in other wards; P ‫؍‬ 0.002). Colonization samples were not predictive for the invasive species in 11/100 cases. Fifty-six percent of the patients had undergone surgery, 51% were intensive care unit (ICU) patients, and 33% had malignant disease. Mortality increased by age (P ‫؍‬ 0.009) and varied by species (36% for C. krusei, 25% for C. parapsilosis, and 14% for other Candida species), severity of underlying disease (47% for ICU patients versus 24% for others; P ‫؍‬ 0.0001), and choice but not timing of initial therapy (12% versus 48% for patients with C. glabrata infection receiving caspofungin versus fluconazole; P ‫؍‬ 0.023). The initial antifungal agent was deemed suboptimal upon species identification in 15% of the cases, which would have been 6.5% if current guidelines had been followed. A large proportion of Danish fungemia patients were severely ill and received suboptimal initial antifungal treatment. Optimization of diagnosis and therapy is possible. Surveillance of fungemia was initiated in Denmark in 2003and has demonstrated a high incidence of this condition and an increasing proportion of isolates belonging to the not fully susceptible species Candida glabrata and C. krusei from a Nordic as well as a global perspective (4, 9-11, 16, 23, 27, 40, 41, 46, 52).A number of recent surveys have provided important information on underlying diseases and host factors in patients with fungemia. The most important factors are (i) critical illness with a prolonged stay in the intensive care unit (ICU); (ii) abdominal surgery, especially if it is complicated or repeated; (iii) low birth weight; (iv) acute necrotizing pancreatitis; (v) malignant disease; (vi) organ transplantation, especially of the liver; (vii) Candida colonization; and (viii) use of antibiotics, central venous catheters, steroids, dialysis, and total parenteral nutrition.The crude 30-day mortality was 30 to 40% in most population-based studies enrolling patients until the turn of the millennium (2,4,12,14,16,29,41,42,49,52) but was lower in recent studies (16,17,36) and higher for ICU pati...

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Section: Methodsmentioning

confidence: 99%

Diagnostic Issues, Clinical Characteristics, and Outcomes for Patients with Fungemia

et al. 2011

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“…using broth microdilution (BMD): that of the Clinical and Laboratory Standards Institute (CLSI) (13,14) and that of the European Committee on Antimicrobial Susceptibility Testing (EUCAST) (23,24,87). These methods provide MIC data for azole and echinocandin antifungal agents that are both quantitatively and qualitatively similar (4,66,71).…”

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Progress in Antifungal Susceptibility Testing of Candida spp. by Use of Clinical and Laboratory Standards Institute Broth Microdilution Methods, 2010 to 2012

2012

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Antifungal susceptibility testing of Candida has been standardized and refined and now may play a useful role in managing Candida infections. Important new developments include validation of 24-h reading times for all antifungal agents and the establishment of species-specific epidemiological cutoff values (ECVs) for the systemically active antifungal agents and both common and uncommon species of Candida . The clinical breakpoints (CBPs) for fluconazole, voriconazole, and the echinocandins have been revised to provide species-specific interpretive criteria for the six most common species. The revised CBPs not only are predictive of clinical outcome but also provide a more sensitive means of identifying those strains with acquired or mutational resistance mechanisms. This brief review serves as an update on the new developments in the antifungal susceptibility testing of Candida spp. using Clinical and Laboratory Standards Institute (CLSI) broth microdilution (BMD) methods.

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“…Consequently, the Clinical and Laboratory Standards Institute (CLSI) and the European Committee on Antimicrobial Susceptibility Testing (EUCAST) have established reference test methods for susceptibility to the major classes of antifungals, which include the polyenes, azoles, and the recently developed echinocandins (1,2). These standards utilize broth macrodilution or microdilution assays for testing yeasts (M27-A3 [3] and E.DEF 7.1 [4]) and filamentous fungi (M38-A2 [5] and E.DEF 9.1 [6]). Procedures, including inoculum preparation, media, assay duration, and endpoint definitions, have now been established for testing some of the more commonly encountered yeast-form pathogenic fungi (i.e., Candida species and Cryptococcus).…”

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Quantitative Microplate-Based Growth Assay for Determination of Antifungal Susceptibility of Histoplasma capsulatum Yeasts

2015

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bStandardized methodologies for determining the antifungal susceptibility of fungal pathogens is central to the clinical management of invasive fungal disease. Yeast-form fungi can be tested using broth macrodilution and microdilution assays. Reference procedures exist for Candida species and Cryptococcus yeasts; however, no standardized methods have been developed for testing the antifungal susceptibility of yeast forms of the dimorphic systemic fungal pathogens. For the dimorphic fungal pathogen Histoplasma capsulatum, susceptibility to echinocandins differs for the yeast and the filamentous forms, which highlights the need to employ Histoplasma yeasts, not hyphae, in antifungal susceptibility tests. To address this, we developed and optimized methodology for the 96-well microtiter plate-based measurement of Histoplasma yeast growth in vitro. Using optical density, the assay is quantitative for fungal growth with a dynamic range greater than 30-fold. Concentration and assay reaction time parameters were also optimized for colorimetric (MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] reduction) and fluorescent (resazurin reduction) indicators of fungal vitality. We employed this microtiter-based assay to determine the antifungal susceptibility patterns of multiple clinical isolates of Histoplasma representing different phylogenetic groups. This methodology fulfills a critical need for the ability to monitor the effectiveness of antifungals on Histoplasma yeasts, the morphological form present in mammalian hosts and, thus, the form most relevant to disease.T he increasing incidence of fungal disease necessitates adequate and timely assessment of antifungal susceptibility to guide the selection and implementation of antifungal therapies. Consequently, the Clinical and Laboratory Standards Institute (CLSI) and the European Committee on Antimicrobial Susceptibility Testing (EUCAST) have established reference test methods for susceptibility to the major classes of antifungals, which include the polyenes, azoles, and the recently developed echinocandins (1, 2). These standards utilize broth macrodilution or microdilution assays for testing yeasts (M27-A3 [3] and E.DEF 7.1 [4]) and filamentous fungi (M38-A2 [5] and E.DEF 9.1 [6]). Procedures, including inoculum preparation, media, assay duration, and endpoint definitions, have now been established for testing some of the more commonly encountered yeast-form pathogenic fungi (i.e., Candida species and Cryptococcus). However, notably missing from the M27-A3 and E.DEF 7.1 standards for testing of yeasts are methods for testing the yeast forms of the dimorphic fungi.The dimorphic fungi that cause systemic disease are characterized by distinct yeast and filamentous morphological states (7,8). Temperature is the principal morphology-determining factor, with yeast forms characterizing infections in mammals (9). Histoplasma capsulatum is the most common clinically encountered dimorphic fungal pathogen in the United States, with an estimated 10,000 to 20,000...

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EUCAST Definitive Document EDef 7.1: method for the determination of broth dilution MICs of antifungal agents for fermentative yeasts

Cited by 464 publications

References 14 publications

Diagnostic Issues, Clinical Characteristics, and Outcomes for Patients with Fungemia

Diagnostic Issues, Clinical Characteristics, and Outcomes for Patients with Fungemia

Progress in Antifungal Susceptibility Testing of Candida spp. by Use of Clinical and Laboratory Standards Institute Broth Microdilution Methods, 2010 to 2012

Quantitative Microplate-Based Growth Assay for Determination of Antifungal Susceptibility of Histoplasma capsulatum Yeasts

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