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Broth Microdilution In Vitro Screening: An Easy and Fast Method to Detect New Antifungal Compounds
Published on: February 14, 2018
[Comparison of broth microdilution and E-test methods for the antifungal susceptibility testing of Candida spp.
Sema Keçeli Ozcan1, Birsen Mutlu, Devrim Dündar
1Kocaeli Universitesi Tip Fakültesi, Tibbi Mikrobiyoloji Anabilim Dali, Kocaeli. keceliozcan@yahoo.com
Abstract:
The incidence of serious fungal infections, particularly invasive Candida infections exhibit an increasing trend in the last decades since the number of patients receiving immunosuppressive treatment is increasing. This situation eventually results in an increment in resistance to antifungal agents. The aim of this study was to compare the standard broth microdilution (BMD) and E-test methods for antifungal susceptibility testing of Candida species isolated from blood cultures in our hospital, against fluconazole, voriconazole, caspofungin and amphotericin B. A total of 46 Candida strains isolated from the blood cultures by BACTEC 9000 (Becton Dickinson, USA) and identified by conventional techniques and API 20C AUX (BioMerieux, France) during January 2006-December 2007, were included into this study. The identification results of the isolates were as follows: C. albicans (23), C. parapsilosis (10), C. tropicalis (5), C. krusei (3), C. famata (2), C. glabrata (1), C. guilliermondii (1), C. kefyr (1). The antifungal susceptibilities were determined by BMD method described in Clinical and Laboratory Standards Institute M27-A3 document and E-test. Only two isolates (C. albicans and C. globrata) were found to be resistant to fluconazole with E-test but susceptible with BMD. The minimal inhibitory concentration (MIC) values of caspofungin were higher (MIC = 1-2 microg/ml) in C. parapsilosis compared to other Candida species using E-test. Only one C. albicans was resistant to voriconazole by E-test (MIC = 4 microg/ml), but it was susceptible by BMD (MIC = 0.08 microg/ml). Since definite resistance breakpoints do not yet exist for amphotericin B, MIC values were considered for amphotericin B and it was found that all strains had identical low MIC values (< 0.002-0.5). When E-test results were compared with the standard BMD results, MIC values were in agreement 80.4% for fluconazole, 84.7% for amphotericin B, 95.6% for voriconazole and 93.4% for caspofungin. These results indicated that the most frequently isolated Candida species among blood cultures was C. albicans, followed by C. parapsilosis and these isolates had low antifungal resistance rates. When voriconazol and caspofungin susceptibilities were considered, both E-test and BMD susceptibility results were in good aggreement in comparison to fluconazol and amphotericin B. E-test can be considered as a compatible method for the antifungal susceptibility testing of Candida species compared to standard broth microdilution method.
Insights
Serious fungal infections are rising, leading to increased antifungal resistance. This study compared standard broth microdilution (BMD) and E-test methods for Candida susceptibility testing, finding E-test to be a compatible alternative to BMD.
Area of Science:
- Clinical Microbiology
- Infectious Diseases
- Mycology
Background:
- Increasing incidence of serious fungal infections, particularly invasive Candida species, due to rising numbers of immunocompromised patients.
- Growing concern over antifungal resistance necessitates accurate and reliable susceptibility testing methods.
- Need for comparative analysis of established and alternative antifungal susceptibility testing techniques.
Purpose of the Study:
- To compare the standard broth microdilution (BMD) method with the E-test for antifungal susceptibility testing of Candida species.
- To evaluate the agreement between BMD and E-test for fluconazole, voriconazole, caspofungin, and amphotericin B.
- To assess the antifungal resistance patterns of Candida species isolated from blood cultures.
Main Methods:
- Collected 46 Candida strains from blood cultures between January 2006 and December 2007.
- Identified Candida species using conventional techniques and API 20C AUX.
- Determined antifungal susceptibility using the Clinical and Laboratory Standards Institute M27-A3 BMD method and E-test.
Main Results:
- High agreement between E-test and BMD: 95.6% for voriconazole, 93.4% for caspofungin, 84.7% for amphotericin B, and 80.4% for fluconazole.
- Candida albicans was the most frequent isolate (23 strains), followed by C. parapsilosis (10 strains), with low antifungal resistance rates.
- Minor discrepancies observed for fluconazole and voriconazole, with E-test showing resistance in two isolates where BMD indicated susceptibility.
Conclusions:
- E-test demonstrates good agreement with the standard BMD method for antifungal susceptibility testing of Candida species.
- Voriconazole and caspofungin showed higher agreement between E-test and BMD compared to fluconazole and amphotericin B.
- E-test can be considered a compatible and reliable method for routine antifungal susceptibility testing of Candida blood isolates.
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