A reversed-phase high-performance liquid chromatographic (HPLC) method, with internal standard quantification, is described for the analysis of itraconazole in human serum. No interference was encountered from over 60 drugs tested. The standard curve was linear from 10 to 10,000 micrograms/l. The detection limit of the method was 10 micrograms/l, with coefficients of variation from 2.2 to 7.8% over a range of itraconazole concentrations from 20 to 1600 micrograms/l. An agar diffusion method is also described with a lowest reproducible limit of 100 micrograms/l. This method had coefficients of variation from 11.0 to 17.1% over a range of itraconazole concentrations from 100 to 1600 micrograms/l. Comparison of the methods showed that HPLC gave much lower values of itraconazole concentrations in patient serum samples than did the microbiological method.
Methotrexate, at concentrations corresponding to drug levels found in vivo, caused a marked reduction in blastospore growth of Candida albicans strains in vitro, but did not affect germ tube elongation during the first 6 h after blastospores were shifted to conditions favouring hyphal formation. Combinations of methotrexate with amphotericin B, flucytosine and itraconazole showed no potentiation or antagonism of the antifungal effect of either component in tests with the blastospore form of C. albicans. In contrast, potentiation occurred when methotrexate-itraconazole combinations were tested for their effect on germ tube elongation of an azole-sensitive strain of C. albicans. This effect was not seen in tests with an azole-resistant strain.
A screening test has been developed for the presumptive identification of Torulopsis (Candida) glabrata from other common clinical isolates of yeast-like fungi. An interlaboratory comparison of a protocol consisting of morphology on cornmeal Tween 80 agar and trehalose fermentation at 42؇C was successful in differentiating T. glabrata from other taxa that are frequent or possible clinical isolates. The screening results for 517 clinical yeast isolates, 241 of which were T. glabrata, were compared with their final identification via commercial systems (API 20C Yeast Identification System [bioMERIEUX, Hazelwood, Mo.] and Rapid Yeast Identification Panel [Dade Microscan, Sacramento, Calif.]). The trehalose screening test has a sensitivity and a specificity of 97.8 and 95.8%, respectively, and a positive predictive value of 97.4% and a negative predictive value of 96.5%. Overall, the trehalose screen had an efficiency rating of 93.9% for ruling in or out T. glabrata. Since T. glabrata represents a substantial part of the workload in a clinical laboratory, a significant reduction in direct and indirect costs should be realized.
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