Comparison of three methods of antifungal susceptibility testing with the proposed NCCLS standard broth macrodilution assay: lack of effect of phenol red. National Committee for Clinical Laboratory Standards. 1995

A M Sugar, and X Liu
Evans Memorial Department of Clinical Research, Department of Medicine, Boston University Medical Center Hospital, Massachusetts 02160, USA.

Three microtiter plate adaptations of the NCCLS proposed standard for antifungal susceptibility testing were evaluated and compared to the NCCLS broth macrodilution method. Thirteen different fungi, including yeasts and moulds were studied. The first microtiter based method was performed exactly as described for the tube dilution assay, with the exception of performance of the assay in 100 microliter in wells of the microtiter plate. The second assay was the same as the first, except for the deletion of phenol red from the RPMI 1640. The third microtiter assay was based on the reduction of the formazan dye, XTT, after only 24 hours of incubation. All three microtiter methods compared favorably with the macrodilution method, when visually read after either 24 or 48 hours of incubation. Minimum inhibitory concentrations obtained by the XTT assay were usually lower than those obtained by the methods requiring a visual end point determination. Results were reproducible and comparable to those obtained with the NCCLS method. We conclude that microtiter plate adaptation of the NCCLS proposed standard is feasible and the presence of phenol red does not alter the results with the drugs tested. A 24 hour assay using XTT may provide a quicker and more quantitative method of susceptibility testing of fungi. Further investigation of this approach is warranted.

UI MeSH Term Description Entries
D008826 Microbial Sensitivity Tests Any tests that demonstrate the relative efficacy of different chemotherapeutic agents against specific microorganisms (i.e., bacteria, fungi, viruses). Bacterial Sensitivity Tests,Drug Sensitivity Assay, Microbial,Minimum Inhibitory Concentration,Antibacterial Susceptibility Breakpoint Determination,Antibiogram,Antimicrobial Susceptibility Breakpoint Determination,Bacterial Sensitivity Test,Breakpoint Determination, Antibacterial Susceptibility,Breakpoint Determination, Antimicrobial Susceptibility,Fungal Drug Sensitivity Tests,Fungus Drug Sensitivity Tests,Sensitivity Test, Bacterial,Sensitivity Tests, Bacterial,Test, Bacterial Sensitivity,Tests, Bacterial Sensitivity,Viral Drug Sensitivity Tests,Virus Drug Sensitivity Tests,Antibiograms,Concentration, Minimum Inhibitory,Concentrations, Minimum Inhibitory,Inhibitory Concentration, Minimum,Inhibitory Concentrations, Minimum,Microbial Sensitivity Test,Minimum Inhibitory Concentrations,Sensitivity Test, Microbial,Sensitivity Tests, Microbial,Test, Microbial Sensitivity,Tests, Microbial Sensitivity
D010637 Phenolsulfonphthalein Red dye, pH indicator, and diagnostic aid for determination of renal function. It is used also for studies of the gastrointestinal and other systems. Phenol Red,Sulfonphthal,Sulphental,Sulphonthal
D005658 Fungi A kingdom of eukaryotic, heterotrophic organisms that live parasitically as saprobes, including MUSHROOMS; YEASTS; smuts, molds, etc. They reproduce either sexually or asexually, and have life cycles that range from simple to complex. Filamentous fungi, commonly known as molds, refer to those that grow as multicellular colonies. Fungi, Filamentous,Molds,Filamentous Fungi,Filamentous Fungus,Fungus,Fungus, Filamentous,Mold
D013778 Tetrazolium Salts Quaternary salts derived from tetrazoles. They are used in tests to distinguish between reducing sugars and simple aldehydes, for detection of dehydrogenase in tissues, cells, and bacteria, for determination of corticosteroids, and in color photography. (From Mall's Dictionary of Chemistry, 5th ed, p455) Tetrazolium Salt,Salt, Tetrazolium,Salts, Tetrazolium

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