Espinel-Ingroff A, Kish C W, Kerkering T M, Fromtling R A, Bartizal K, Galgiani J N, Villareal K, Pfaller M A, Gerarden T, Rinaldi M G
Medical College of Virginia, Richmond 23298-0049.
J Clin Microbiol. 1992 Dec;30(12):3138-45. doi: 10.1128/jcm.30.12.3138-3145.1992.
A collaborative comparison of macro- and microdilution antifungal susceptibility tests was performed in five laboratories. MICs of amphotericin B, fluconazole, flucytosine, and ketoconazole were determined in all five centers against 95 coded isolates of Candida spp., Cryptococcus neoformans, and Torulopsis glabrata. A standard protocol with the following National Committee for Clinical Laboratory Standards Subcommittee on Antifungal Susceptibility Testing recommendations was used: an inoculum standardized by spectrophotometer, buffered (RPMI 1640) medium (pH 7.0), incubation at 35 degrees C, and an additive drug dilution procedure. Two inoculum sizes were tested (1 x 10(4) to 5 x 10(3) to 2.5 x 10(3) CFU/ml) and three scoring criteria were evaluated for MIC endpoint determinations, which were scored as 0 (optically clear), < or = 1 (slightly hazy turbidity), and < or = 2 (prominent decrease in turbidity compared with that of the growth control). Overall intra- and interlaboratory reproducibility was optimal with the low-density inoculum, the second-day readings, and MICs scored as either 1 or 2. The microdilution MICs demonstrated interlaboratory agreement with most of the four drugs higher than or similar to that of the macrodilution MICs. In general, there was good interlaboratory agreement with amphotericin B, fluconazole, and flucytosine; ketoconazole gave more variable results.
五个实验室对宏观和微观稀释抗真菌药敏试验进行了协作比较。在所有五个中心测定了两性霉素B、氟康唑、氟胞嘧啶和酮康唑针对95株编码的念珠菌属、新型隐球菌和光滑球拟酵母菌的最低抑菌浓度(MIC)。采用了具有以下美国国家临床实验室标准委员会抗真菌药敏试验小组委员会建议的标准方案:用分光光度计标准化接种物、缓冲(RPMI 1640)培养基(pH 7.0)、在35摄氏度孵育以及附加药物稀释程序。测试了两种接种物浓度(1×10⁴至5×10³至2.5×10³CFU/ml),并评估了三种用于MIC终点判定的评分标准,分别评分为0(光学上澄清)、≤1(轻微浑浊)和≤2(与生长对照相比浊度显著降低)。总体而言,实验室内部和实验室间的重复性在低密度接种物、第二天读数以及评分为1或2的MIC时最佳。微观稀释法测定的MIC显示,四种药物中大多数的实验室间一致性高于或类似于宏观稀释法测定的MIC。一般来说,两性霉素B、氟康唑和氟胞嘧啶的实验室间一致性良好;酮康唑的结果变异性更大。