scholarly journals Comparative Evaluation of NCCLS M27-A and EUCAST Broth Microdilution Procedures for Antifungal Susceptibility Testing of Candida Species

2002 ◽  
Vol 46 (11) ◽  
pp. 3644-3647 ◽  
Author(s):  
Manuel Cuenca-Estrella ◽  
Wendy Lee-Yang ◽  
Meral A. Ciblak ◽  
Beth A. Arthington-Skaggs ◽  
Emilia Mellado ◽  
...  
Keyword(s):  
Amphotericin B ◽  
Correlation Coefficient ◽  
Intraclass Correlation Coefficient ◽  
Laboratory Study ◽  
Candida Species ◽  
Comparative Evaluation ◽  
Susceptibility Testing ◽  
Antifungal Susceptibility ◽  
Intraclass Correlation ◽  
Broth Microdilution

ABSTRACT A two-laboratory study was performed to evaluate the correlation between the NCCLS M27-A and EUCAST microdilution procedures for antifungal testing of Candida spp. A panel of 109 bloodstream isolates was tested against amphotericin B, flucytosine, fluconazole, and itraconazole. Overall, the agreement was 92% and the intraclass correlation coefficient was 0.90 (P < 0.05).

scholarly journals Comparative Evaluation of a New Commercial Colorimetric Microdilution Assay (SensiQuattro Candida EU) with MIC Test Strip and EUCAST Broth Microdilution Methods for Susceptibility Testing of Invasive Candida Isolates

2014 ◽  
Vol 53 (1) ◽  
pp. 255-261 ◽  
Author(s):  
Hedda Luise Koehling ◽  
Birgit Willinger ◽  
Jan Buer ◽  
Peter-Michael Rath ◽  
Joerg Steinmann
Keyword(s):  
Amphotericin B ◽  
Comparative Evaluation ◽  
Antifungal Agents ◽  
Susceptibility Testing ◽  
Antifungal Susceptibility ◽  
Test Strip ◽  
Broth Microdilution ◽  
Content Type ◽  
Immunosuppressed Patients ◽  
Level Of Agreement

Candidemia is an important cause of morbidity and mortality in immunosuppressed patients.Candidaisolates must be cultivated, identified, and tested for susceptibility. We compared the performance of a new colorimetric broth microdilution panel (SensiQuattroCandidaEU) for antifungal susceptibility testing to that of Liofilchem's MIC test strip and the EUCAST reference broth microdilution protocol. We tested 187 blood culture isolates of 5Candidaspp. (120C. albicans, 38C. glabrata, 10C. parapsilosis, 12C. tropicalis, and 7C. krusei) against seven antifungal agents (amphotericin B, fluconazole, voriconazole, posaconazole, caspofungin, anidulafungin, and micafungin) and interpreted the MICs according to the EUCAST recommendations. If applicable, the overall essential agreement (EA) of the SensiQuattro panel with the reference broth microdilution was slightly higher forC. albicans(87%) than for other species (85.8%). We found that SensiQuattro performed best in testing amphotericin B (EA, 100%), voriconazole (EA, 93.7%), and posaconazole (EA, 94.8%) againstC. albicans, but its error rate for this species was high (29.6%) because of mainly major errors (26.7%) in testing anidulafungin and micafungin. Compared to the SensiQuattro panel, the MIC test strip exhibited a higher level of agreement for most isolates. SensiQuattro assays are easy to perform, but they are currently not suitable for testing echinocandins againstCandidaspp.

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 Comparison of the Sensititre YeastOne and CLSI M38-A2 Microdilution Methods in Determining the Activity of Amphotericin B, Itraconazole, Voriconazole, and Posaconazole against Aspergillus Species

2018 ◽  
Vol 56 (10) ◽  
Author(s):  
Hsuan-Chen Wang ◽  
Ming-I Hsieh ◽  
Pui-Ching Choi ◽  
Chi-Jung Wu
Keyword(s):  
Aspergillus Fumigatus ◽  
Amphotericin B ◽  
Antifungal Agents ◽  
Susceptibility Testing ◽  
Geometric Mean ◽  
Antifungal Susceptibility ◽  
Reference Method ◽  
Aspergillus Species ◽  
Broth Microdilution ◽  
Content Type

ABSTRACT This study compared the YeastOne and reference CLSI M38-A2 broth microdilution methods for antifungal susceptibility testing of Aspergillus species. The MICs of antifungal agents were determined for 100 Aspergillus isolates, including 54 Aspergillus fumigatus (24 TR34/L98H isolates), 23 A. flavus, 13 A. terreus, and 10 A. niger isolates. The overall agreement (within 2 2-fold dilutions) between the two methods was 100%, 95%, 92%, and 90% for voriconazole, posaconazole, itraconazole, and amphotericin B, respectively. The voriconazole geometric mean (GM) MICs were nearly identical for all isolates using both methods, whereas the itraconazole and posaconazole GM MICs obtained using the YeastOne method were approximately 1 dilution lower than those obtained using the reference method. In contrast, the amphotericin B GM MIC obtained using the YeastOne method was 3.3-fold higher than that observed using the reference method. For the 24 A. fumigatus TR34/L98H isolates assayed, the categorical agreement (classified according to the CLSI epidemiological cutoff values) was 100%, 87.5%, and 83.3% for itraconazole, voriconazole, and posaconazole, respectively. For four A. niger isolates, the itraconazole MICs were >8 μg/ml using the M38-A2 method due to trailing growth, whereas the corresponding itraconazole MICs obtained using the YeastOne method were all ≤0.25 μg/ml without trailing growth. These data suggest that the YeastOne method can be used as an alternative for azole susceptibility testing of Aspergillus species and for detecting the A. fumigatus TR34/L98H isolates but that this method fails to detect A. niger isolates exhibiting trailing growth with itraconazole. Additionally, for isolates with azole MICs that approach or that are at susceptibility breakpoints or with high amphotericin B MICs detected using the YeastOne method, further MIC confirmation using the reference CLSI method is needed.

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

scholarly journals Comparison of the EUCAST and CLSI Broth Microdilution Methods for Testing Isavuconazole, Posaconazole, and Amphotericin B against Molecularly Identified Mucorales Species

2015 ◽  
Vol 59 (12) ◽  
pp. 7882-7887 ◽  
Author(s):  
Anuradha Chowdhary ◽  
Pradeep Kumar Singh ◽  
Shallu Kathuria ◽  
Ferry Hagen ◽  
Jacques F. Meis
Keyword(s):  
Amphotericin B ◽  
Susceptibility Testing ◽  
Antifungal Susceptibility ◽  
Antifungal Susceptibility Testing ◽  
Broth Microdilution ◽  
Content Type

ABSTRACTWe compared EUCAST and CLSI antifungal susceptibility testing (AFST) methods for triazoles and amphotericin B against 124 clinicalMucoralesisolates. The EUCAST method yielded MIC values 1- to 3-fold dilutions higher than those of the CLSI method for amphotericin B. The essential agreements between the two methods for triazoles were high, i.e., 99.1% (voriconazole), 98.3% (isavuconazole), and 87% (posaconazole), whereas it was significantly lower for amphotericin B (66.1%). Strategies for harmonization of the two methods forMucoralesAFST are warranted.

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