In vitro anti fungal efficacy test
Italy Pharmaceutical Contract Research Organization needed for In vitro efficacy test on an antifungal compound. The test is a reproduction of a previous test that whas publicized in Journal of Med Chem, details can be obtained upon request. The strains that need to be tested against are candida strains, including some azole resistant strains, more specifically Candida albicans, Candida parapsilosis, Candida krusei, and Candida tropicalis.
Reproduction of a previous test in which:
The EUCAST susceptibility testing protocol, including the update for testing of non-fermenting yeasts (Cryptococcus spp. and Geotrichum spp.), was used for the determination of the minimal inhibitory concentrations (MICs) of antifungal agents. Results were read after 24, 48, and 72 h of incubation at 37 °C. The MIC was defined as the lowest compound concentration which prevented visible growth.
Aliquots of 10 ?L were removed from the cells corresponding to the MIC, 2MIC, 4MIC, and 8MIC and serially 10-fold diluted in sterile saline, to minimize the drug carry-over. An aliquot of 10 ?L of each dilution was plated on Sabouraud dextrose agar plates. The plates were incubated at 37 °C for 24/48 h, and the number of CFUs was counted. For Cryptococcus spp. and Geotrichum spp., incubation was for 48 h to facilitate colony counting.28 When less than 100 CFU/mL was expected, 10 ?L of sample was plated without dilution.
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