Multicenter evaluation of the reproducibility of the proposed antifungal susceptibility testing method for fermentative yeasts of the Antifungal Susceptibility Testing Subcommittee of the European Committee on Antimicrobial Susceptibility Testing (AFST-EUCAST)
Multicenter evaluation of the reproducibility of the proposed antifungal susceptibility testing method for fermentative yeasts of the Antifungal Susceptibility Testing Subcommittee of the European Committee on Antimicrobial Susceptibility Testing (AFST-EUCAST)
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DOI：
10.1046/j.1469-0691.2003.00592.x
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发表时间：
2003-06-01
影响因子：
14.2
通讯作者：
Rodríguez-Tudela, JL
中科院分区：
文献类型：
--
作者：
Cuenca-Estrella, M;Moore, CB;Rodríguez-Tudela, JL
目的评价发酵酵母菌药敏试验新标准的实验室内和实验室间的重复性。该标准基于美国国家临床实验室标准委员会(NCCLS)的M27-A程序，但加入了几项修改，包括分光光度法生长相关终点读数。对6种念珠菌(白色念珠菌、热带念珠菌、近平假丝酵母菌、光滑念珠菌、克鲁斯念珠菌、卢西坦念珠菌各1株)和2株质控菌株(克鲁斯念珠菌ATCC6258和近平假丝酵母菌ATCC22019)进行了检测。在RPMI-2%葡萄糖，pH 7.0的情况下，以微量滴定的形式在三天内进行三重试验。对氟胞嘧啶、氟康唑和伊曲康唑进行了检测。总共有3888个MIC值被纳入分析。结果2 4h和48h组内一致性分别为99%和96%，ICC值分别为0 98和0 97(P<；0 0 5)。两株具有拖尾效应的菌株在48h的室内符合率为92%，ICCs为0.91，24 h和48h的室内符合率分别为94%和88%，ICCs为0.93和0.91(P<；0.05)。培养48h后出现拖尾的菌株，其一致性和ICCs值较低。伊曲康唑的重复性最低。结论EUCAST抗真菌药敏试验新方法是一种实验室内和实验室间可重复使用的方法，与NCCLS批准的方法相比具有许多优点。
Objective To evaluate the intra- and inter-laboratory reproducibility of a new standard for susceptibility testing of fermentative yeasts. This standard is based on the M27-A procedure of the National Committee for Clinical Laboratory Standards (NCCLS), but incorporates several modifications, including spectrophotometric growth-dependent endpoint reading.Methods Nine laboratories participated in the study. Common material lots were used to test six Candida species (one each of C. albicans, C. tropicalis, C. parapsilosis, C. glabrata, C. krusei, and C. lusitaniae), and two quality control strains (C. krusei ATCC6258 and C. parapsilosis ATCC22019). Triplicate testing on three separate days was performed in microtiter format with RPMI-2% glucose, pH 7.0. Flucytosine, fluconazole and itraconazole were tested. In total, 3888 MIC values were included in the analyses. Reproducibility was calculated by means of agreement (percentage of MICs within one two-fold dilution of the mode) and intraclass correlation coefficient (ICC, maximum value of 1).Results The average intra-laboratory agreements were 99% and 96% after 24 h and 48 h of incubation, respectively, with ICCs of 0.98 and 0.97 (P < 0.05). Two strains exhibiting a trailing effect showed intra-laboratory agreement of 92% and ICCs of < 0.91 at 48 h. The inter-laboratory agreement was 94% and 88% after 24 h and 48 h, respectively, with ICCs of 0.93 and 0.91 (P < 0.05). Lower values of agreement and ICCs were obtained for strains exhibiting trailing after 48 h of incubation. Itraconazole yielded the lowest values of reproducibility.Conclusion The new procedure of EUCAST for antifungal susceptibility testing is a reproducible method within and between laboratories and offers several advantages over the NCCLS approved method.