How to: EUCAST recommendations on the screening procedure E.Def 10.1 for the detection of azole resistance in Aspergillus fumigatus isolates using four-well azole-containing agar plates

Guinea, Jesús; Verweij, Paul E.; Meletiadis, Joseph; Mouton, Johan W.; Barchiesi, Francesco; Arendrup, Maiken Cavling; Arıkan-Akdağlı, Sevtap; Castanheira, Mariana; Chryssanthou, Erja; Friberg, Nathalie; Järv, H.; Климко, Н Н; Kurzai, Oliver; Lagrou, Katrien; Lass‐Flörl, Cornelia; Mareș, Mihai; Matos, Tadeja; Moore, Caroline B.; Müehlethaler, Konrad; Rogers, Thomas R.; Andersen, Cecilie Torp; Velegraki, Aristea

doi:10.1016/j.cmi.2018.09.008

Cited by 70 publications

(55 citation statements)

References 21 publications

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“…Its performance is not affected by the observer's experience in mycology . The European Committee on Antimicrobial Susceptibility Testing Subcommittee on Antifungal Susceptibility Testing has released a standard method for performing azole‐containing agar plate tests . The YeastOne method was used for further testing of positive azole resistance screening isolates, to evaluate the MICs of several antifungals.…”

Section: Discussionmentioning

confidence: 99%

Azole resistance in Aspergillus species in Southern Taiwan: An epidemiological surveillance study

Chen

Kuo

Wang

et al. 2019

Mycoses

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Summary Poor clinical outcomes for invasive aspergillosis are associated with antifungal resistance. Performing antifungal susceptibility tests on clinically relevant Aspergillus isolates from patients and environmental regions with known azole resistance is recommended. The aim of the study was to assess the presence of azole resistance in clinical Aspergillus spp. isolates and those from hospital environments and farmlands within a 40 km radius of the hospital. Clinical Aspergillus spp. isolates were cultured, as well as environmental Aspergillus spp. isolates obtained from air samples. Samples were subcultured in azole‐containing agar plates. Isolates with a positive screening test were subjected to YeastOne methods to determine their minimum inhibitory concentrations of antifungals. Resistance mechanisms were investigated in the azole‐resistant Aspergillus spp. isolates. No azole‐resistant clinical or environmental A flavus, A oryaze, A niger or A terreus isolates were found in the present study. All A fumigatus clinical isolates were azole‐susceptible. Seven A fumigatus environmental isolates were associated with cyp51A mutations, including two that harboured TR34/L98H mutations with S297T/F495I substitutions, two with TR34/L98H mutations and three with TR46/Y121F/T289A mutations. One of these isolates was collected from farmland, one was from A ward and five were from B ward. The proportion of azole‐resistant A fumigatus was 10.2% (6/59) and 3.2% (1/31) in the hospital environments and the farmlands near the hospital, respectively. The results showed that azole‐resistant A fumigatus existed within hospital environments. This emphasises the importance of periodic surveillance in hospital environments and monitoring for the emergence of azole‐resistant A fumigatus clinical isolates.

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Section: Discussionmentioning

confidence: 99%

Azole resistance in Aspergillus species in Southern Taiwan: An epidemiological surveillance study

Chen

Kuo

Wang

et al. 2019

Mycoses

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“…Currently, the only method recommended by the guidelines to detect azole resistance in patients suspected of having an infection caused by azole-resistant A. fumigatus is susceptibility testing by culture [6,8,38,39]. However, more than half of BAL cultures are reported with no fungal growth and IA diagnosis is made based on radiology, fungal biomarker and molecular test results which do not provide information on resistance [40][41][42][43].…”

Section: Diagnosis Of Aspergillosismentioning

confidence: 99%

“…On the other hand, even in the case of a positive culture and access to susceptibility testing or resistance screening, the turnaround times for these culture-based methods are long, and the delivery of results in clinically useful time frame is challenging. Particularly in smaller centres, where susceptibility testing is not available on site, this timeline often exceeds one week [39,44].…”

Section: Diagnosis Of Aspergillosismentioning

confidence: 99%

Detecting Azole-Antifungal Resistance in Aspergillus fumigatus by Pyrosequencing

Torre

Frazer

Rautemaa‐Richardson

2020

JoF

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Guidelines on the diagnosis and management of Aspergillus disease recommend a multi-test approach including CT scans, culture, fungal biomarker tests, microscopy and fungal PCR. The first-line treatment of confirmed invasive aspergillosis (IA) consists of drugs in the azole family; however, the emergence of azole-resistant isolates has negatively impacted the management of IA. Failure to detect azole-resistance dramatically increases the mortality rates of azole-treated patients. Despite drug susceptibility tests not being routinely performed currently, we suggest including resistance testing whilst diagnosing Aspergillus disease. Multiple tools, including DNA sequencing, are available to screen for drug-resistant Aspergillus in clinical samples. This is particularly beneficial as a large proportion of IA samples are culture negative, consequently impeding susceptibility testing through conventional methods. Pyrosequencing is a promising in-house DNA sequencing method that can rapidly screen for genetic hotspots associated with antifungal resistance. Pyrosequencing outperforms other susceptibility testing methods due to its fast turnaround time, accurate detection of polymorphisms within critical genes, including simultaneous detection of wild type and mutated sequences, and—most importantly—it is not limited to specific genes nor fungal species. Here we review current diagnostic methods and highlight the potential of pyrosequencing to aid in a diagnosis complete with a resistance profile to improve clinical outcomes.

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“…Demonstrating tissue invasion by hyphae through microscopic examination of biopsy or autopsy material provides a diagnosis of proven invasive fungal infection. However, the sensitivity of microscopy for IA is 50% at best [14,35]. PCR approach can increase the sensitivity to 57-> 90% depending on the amount of fungi in the sample.…”

Section: Microbiological Findingsmentioning

confidence: 99%

Fungal brain infection—no longer a death sentence

et al. 2020

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The aim of this case series was to provide a modern cohort of patients with cerebral aspergillosis and show the effectiveness of modern treatment concepts. In a 10-year period from January 2009 to January 2019, we identified 10 patients (6 male, 4 female) who received surgery or frameless stereotactic drainage of a cerebral aspergilloma at our center. Patients’ and disease characteristics were recorded. The median age was 65 (range 45 to 83). We conducted 133 cranial surgeries in 100 patients due to cerebral brain abscess (BA) during that time, which leads to a percentage of 10% of aspergilloma within BAs in our patient sample. We performed 3.1 surgeries per patient followed by antifungal treatment for 6 months (= median) according to the microbiological findings. Regarding comorbidities, the mean Charlson comorbidity index (CCI) at the time of admission was 5, representing an estimated 10-year survival of 21%. Six (60%) of 10 patients showed conditions of immunosuppression, one suffered endocarditis after replacement of aortic valves. Four patients showed associated frontobasal bone destruction, mycotic aneurysms, or thromboses. The mean duration of hospital stay was 37 days. Mortality was much lower than in literature. Sixty percent of the patients died during the follow-up period. The outcome of the two immunocompetent patients was more favorable. Cerebral aspergillosis is a rare, but still life-threatening, condition, which predominantly occurs in immunosuppressive conditions. Due to radical surgical and antifungal therapy for several months, mortality can be reduced dramatically.

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How to: EUCAST recommendations on the screening procedure E.Def 10.1 for the detection of azole resistance in Aspergillus fumigatus isolates using four-well azole-containing agar plates

Abstract: Implications: This easy-to-use screening procedure for the detection of azole resistance in clinical A. fumigatus isolates will allow rapid testing in the daily routine of the microbiology laboratory and thus facilitate earlier appropriate therapy.

Cited by 70 publications

References 21 publications

Azole resistance in Aspergillus species in Southern Taiwan: An epidemiological surveillance study

Azole resistance in Aspergillus species in Southern Taiwan: An epidemiological surveillance study

Detecting Azole-Antifungal Resistance in Aspergillus fumigatus by Pyrosequencing

Fungal brain infection—no longer a death sentence

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