scholarly journals Updated EUCAST Clinical Breakpoints against Aspergillus, Implications for the Clinical Microbiology Laboratory

2020 ◽  
Vol 6 (4) ◽  
pp. 343
Author(s):  
Jesús Guinea
Keyword(s):  
Amphotericin B ◽  
Susceptibility Testing ◽  
Antifungal Susceptibility ◽  
Common Species ◽  
Antifungal Susceptibility Testing ◽  
Clinical Microbiology Laboratory ◽  
Closely Related Species ◽  
Exposure Category ◽  
Clinical Breakpoints ◽  
Definition Of

Azole resistance poses a problem for the management of patients with invasive aspergillosis. Former species are in fact groups of closely related species (or complexes); cryptic species frequently show high antifungal resistance. The European Committee on Antimicrobial Susceptibility Testing (EUCAST) Definitive Document (E.Def) 9.3.2 includes guidelines for antifungal susceptibility testing on Aspergillus spp. and clinical breakpoints for amphotericin B, itraconazole, voriconazole, posaconazole, and isavuconazole against A. flavus, A. fumigatus, A. nidulans, A. niger, and A. terreus. New clinical breakpoints were released in February 2020 and one of the most relevant modifications was the definition of the new “susceptible, increased exposure” (formerly “intermediate”) category. Another relevant change was the adoption of the concept of area of technical uncertainty (ATU) that refers to problematic areas which involve uncertainty of susceptibility categorisation (e.g., when minimum inhibitory concentrations (MICs) for susceptible and resistant organisms overlap). To accommodate both the new “susceptible, increased exposure” category and the concept of ATU, MICs of azoles and amphotericin B that fall in the former “intermediate” category have been automatically categorized as either R (amphotericin B) or ATU (triazoles). Finally, EUCAST-AFST (Antifungal Susceptibility Testing) decided to adopt new breakpoints for less common species provided that the epidemiological cut-off value (ECOFF) is below or comparable to the breakpoint for the type species (A. fumigatus).

Use of spectrophotometric reading for in vitro antifungal susceptibility testing ofAspergillusspp.

1999 ◽  
Vol 45 (10) ◽  
pp. 871-874 ◽  
Author(s):  
Eric Dannaoui ◽  
Florence Persat ◽  
Marie-France Monier ◽  
Elisabeth Borel ◽  
Marie-Antoinette Piens ◽  
...  
Keyword(s):  
Amphotericin B ◽  
Susceptibility Testing ◽  
Clinical Laboratory ◽  
Antifungal Susceptibility ◽  
Reference Method ◽  
Antifungal Susceptibility Testing ◽  
Aspergillus Species ◽  
National Committee ◽  
Broth Microdilution Method

A comparative study of visual and spectrophotometric MIC endpoint determinations for antifungal susceptibility testing of Aspergillus species was performed. A broth microdilution method adapted from the National Committee for Clinical Laboratory Standards (NCCLS) was used for susceptibility testing of 180 clinical isolates of Aspergillus species against amphotericin B and itraconazole. MICs were determined visually and spectrophotometrically at 490 nm after 24, 48, and 72h of incubation, and MIC pairs were compared. The agreement between the two methods was 99% for amphotericin B and ranged from 95 to 98% for itraconazole. It is concluded that spectrophotometric MIC endpoint determination is a valuable alternative to the visual reference method for susceptibility testing of Aspergillus species.Key words: antifungal, susceptibility testing, Aspergillus, spectrophotometric reading.

scholarly journals Flow Cytometry Antifungal Susceptibility Testing of Pathogenic Yeasts other than Candida albicans and Comparison with the NCCLS Broth Microdilution Test

2000 ◽  
Vol 44 (10) ◽  
pp. 2752-2758 ◽  
Author(s):  
Rama Ramani ◽  
Vishnu Chaturvedi
Keyword(s):  
Flow Cytometry ◽  
Candida Albicans ◽  
Amphotericin B ◽  
Susceptibility Testing ◽  
Antifungal Susceptibility ◽  
Antifungal Susceptibility Testing ◽  
Broth Microdilution ◽  
Nitrogen Base ◽  
Microdilution Method ◽  
Broth Microdilution Method

ABSTRACT Candida species other than Candida albicansfrequently cause nosocomial infections in immunocompromised patients. Some of these pathogens have either variable susceptibility patterns or intrinsic resistance against common azoles. The availability of a rapid and reproducible susceptibility-testing method is likely to help in the selection of an appropriate regimen for therapy. A flow cytometry (FC) method was used in the present study for susceptibility testing ofCandida glabrata, Candida guilliermondii,Candida krusei, Candida lusitaniae,Candida parapsilosis, Candida tropicalis, andCryptococcus neoformans based on accumulation of the DNA binding dye propidium iodide (PI). The results were compared with MIC results obtained for amphotericin B and fluconazole using the NCCLS broth microdilution method (M27-A). For FC, the yeast inoculum was prepared spectrophotometrically, the drugs were diluted in either RPMI 1640 or yeast nitrogen base containing 1% dextrose, and yeast samples and drug dilutions were incubated with amphotericin B and fluconazole, respectively, for 4 to 6 h. Sodium deoxycholate and PI were added at the end of incubation, and fluorescence was measured with a FACScan flow cytometer (Becton Dickinson). The lowest drug concentration that showed a 50% increase in mean channel fluorescence compared to that of the growth control was designated the MIC. All tests were repeated once. The MICs obtained by FC for all yeast isolates except C. lusitaniae were in very good agreement (within 1 dilution) of the results of the NCCLS broth microdilution method. Paired ttest values were not statistically significant (P = 0.377 for amphotericin B; P = 0.383 for fluconazole). Exceptionally, C. lusitaniae isolates showed higher MICs (2 dilutions or more) than in the corresponding NCCLS broth microdilution method for amphotericin B. Overall, FC antifungal susceptibility testing provided rapid, reproducible results that were statistically comparable to those obtained with the NCCLS method.

scholarly journals Comparing Etest and Broth Microdilution for Antifungal Susceptibility Testing of the Most-Relevant Pathogenic Molds

2015 ◽  
Vol 53 (10) ◽  
pp. 3176-3181 ◽  
Author(s):  
Frédéric Lamoth ◽  
Barbara D. Alexander
Keyword(s):  
Amphotericin B ◽  
Antifungal Therapy ◽  
Susceptibility Testing ◽  
Antifungal Susceptibility ◽  
Multidrug Resistant ◽  
Antifungal Susceptibility Testing ◽  
Diffusion Method ◽  
Broth Microdilution ◽  
Content Type ◽  
Broth Microdilution Method

Invasive mold infections are life-threatening diseases for which appropriate antifungal therapy is crucial. Their epidemiology is evolving, with the emergence of triazole-resistantAspergillusspp. and multidrug-resistant non-Aspergillusmolds. Despite the lack of interpretive criteria, antifungal susceptibility testing of molds may be useful in guiding antifungal therapy. The standard broth microdilution method (BMD) is demanding and requires expertise. We assessed the performance of a commercialized gradient diffusion method (Etest method) as an alternative to BMD. The MICs or minimal effective concentrations (MECs) of amphotericin B, voriconazole, posaconazole, caspofungin, and micafungin were assessed for 290 clinical isolates of the most representative pathogenic molds (154Aspergillusand 136 non-Aspergillusisolates) with the BMD and Etest methods. Essential agreements (EAs) within ±2 dilutions of ≥90% between the two methods were considered acceptable. EAs for amphotericin B and voriconazole were >90% for most potentially susceptible species. For posaconazole, the correlation was acceptable forMucoromycotinabut Etest MIC values were consistently lower forAspergillusspp. (EAs of <90%). Excellent EAs were found for echinocandins with highly susceptible (MECs of <0.015 μg/ml) or intrinsically resistant (MECs of >16 μg/ml) strains. However, MEC determinations lacked consistency between methods for strains exhibiting mid-range MECs for echinocandins. We concluded that the Etest method is an appropriate alternative to BMD for antifungal susceptibility testing of molds under specific circumstances, including testing with amphotericin B or triazoles for non-Aspergillusmolds (MucoromycotinaandFusariumspp.). Additional study of molecularly characterized triazole-resistantAspergillusisolates is required to confirm the ability of the Etest method to detect voriconazole and posaconazole resistance amongAspergillusspp.

scholarly journals Etest Cannot Be Recommended forIn VitroSusceptibility Testing of Mucorales

2015 ◽  
Vol 59 (6) ◽  
pp. 3663-3665 ◽  
Author(s):  
Rita Caramalho ◽  
Elisabeth Maurer ◽  
Ulrike Binder ◽  
Ricardo Araújo ◽  
Somayeh Dolatabadi ◽  
...  
Keyword(s):  
Amphotericin B ◽  
Antimicrobial Susceptibility ◽  
Active Drug ◽  
Susceptibility Testing ◽  
Antifungal Susceptibility ◽  
Antifungal Susceptibility Testing ◽  
Broth Microdilution ◽  
European Committee ◽  
Susceptibility Patterns

ABSTRACTAmphotericin B and posaconazole susceptibility patterns were determined for the most prevalent Mucorales, following EUCAST (European Committee on Antimicrobial Susceptibility Testing) broth microdilution guidelines. In parallel, Etest was performed and evaluated against EUCAST. The overall agreement of MICs gained with Etest and EUCAST was 75.1%; therefore, Etest cannot be recommended for antifungal susceptibility testing of Mucorales. Amphotericin B was the most active drug against Mucorales speciesin vitro, while the activities of posaconazole were more restricted.

scholarly journals Disk diffusion Method in Enriched Mueller Hinton agar for determining susceptibility of Candida isolates from various clinical specimens

2020 ◽  
Vol 28 (1) ◽  
pp. 28-33
Author(s):  
Nabeela Mahboob ◽  
Hasina Iqbal ◽  
Mushtaque Ahmed ◽  
Md Mehedi Hasan Magnet ◽  
Kazi Zulfiquer Mamun
Keyword(s):  
Methylene Blue ◽  
Amphotericin B ◽  
Candida Species ◽  
Susceptibility Testing ◽  
Antifungal Susceptibility ◽  
Antifungal Susceptibility Testing ◽  
Disk Diffusion ◽  
Clinical Specimens ◽  
Mueller Hinton ◽  
Mueller Hinton Agar

Background: Candida species are responsible for various clinical infections ranging from mucocutaneous infection to life threatening invasive diseases. Recently there is a serious concern with increased resistance of antifungal drugs and its consequences. Thus, identification of Candida and its antifungal susceptibility testing has a paramount significance in the management of Candidal infections. The aim of the study was to determine antifungal susceptibility pattern of Candida by Mueller-Hinton agar media supplemented with glucose and methylene blue for disk diffusion testing of fluconazole, miconazole, clotrimazole, amphotericin B and nystatin. Methods: A total of 35 Candida species was isolated from 2000 clinical specimens over 6 month’s period from July 2016 to December 2016. Growths on Blood agar and chromogenic agar were evaluated for colony appearance and microscopic examination. Antifungal susceptibility testing was performed by disk diffusion using Mueller-Hinton agar supplemented with glucose and methylene blue. Results: Candida species were more sensitive to clotrimazole (88.58%) and amphotericin B (88.58%) followed by nystatin ((77.14%), miconazole (74.29%) whereas fluconazole showed the highest level of resistance (60%). Conclusions: The increase in resistance to fluconazole is of serious concern as it is the most commonly used azole for candidiasis. The sensitivity profile of Candida isolates will be helpful to choose appropriate antifungal agents, thus decreasing patient’s morbidity and mortality. J Dhaka Medical College, Vol. 28, No.1, April, 2019, Page 28-33

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