Clancy C J, Nguyen M H
Department of Medicine, College of Medicine, University of Florida, and the Veterans Affairs Medical Center, Gainesville 32610, USA.
J Clin Microbiol. 1997 Nov;35(11):2878-82. doi: 10.1128/jcm.35.11.2878-2882.1997.
We determined the fluconazole MICs for 101 clinical isolates of Candida and Cryptococcus neoformans using the macro- and microdilution methods recommended by the National Committee for Clinical Laboratory Standards. We compared the MICs obtained by these methods with those obtained by a photometric assay that quantified the reduction of 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) by viable fungi. The MIC determined by this method was defined as the highest fluconazole concentration associated with the first precipitous drop in optical density. For Candida, both the MTT and the microdilution methods demonstrated excellent agreement with the standard macrodilution method. The MTT method, however, generated MICs at 24 h that were comparable to those generated by the standard macrodilution method, whereas the microdilution method required 48 h. For C. neoformans, the levels of agreement between the MICs determined by the MTT and microdilution methods after 48 h and those determined by the standard 72-h macrodilution method were 94% (29 of 31) and 94% (29 of 31), respectively. The MTT method therefore provided results comparable to those of currently recommended methods and had the advantages of a more rapid turnaround time and potential adaptability to use as an automated system. Furthermore, the MICs determined by the MTT method were determined photometrically, thereby eliminating reader bias.
我们使用美国国家临床实验室标准委员会推荐的常量和微量稀释法,测定了101株白色念珠菌和新型隐球菌临床分离株的氟康唑最低抑菌浓度(MIC)。我们将这些方法获得的MIC与通过光度法获得的MIC进行了比较,该光度法通过活菌真菌对3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2H-四氮唑溴盐(MTT)的还原进行定量。通过该方法测定的MIC定义为与光密度首次急剧下降相关的最高氟康唑浓度。对于白色念珠菌,MTT法和微量稀释法与标准常量稀释法均显示出极好的一致性。然而,MTT法在24小时产生的MIC与标准常量稀释法产生的MIC相当,而微量稀释法则需要48小时。对于新型隐球菌,MTT法和微量稀释法在48小时后测定的MIC与标准72小时常量稀释法测定的MIC之间的一致水平分别为94%(31株中的29株)和94%(31株中的29株)。因此,MTT法提供的结果与目前推荐的方法相当,并且具有周转时间更快以及有可能适用于自动化系统的优点。此外,MTT法测定的MIC是通过光度法测定的,从而消除了读数偏差。