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米罗那特 AM 与 CLSI 肉汤微量稀释法检测四种常用抗真菌药物对曲霉菌属体外药敏试验的比较评估

Comparative Evaluation of MIRONAUT-AM and CLSI broth microdilution method for antifungal susceptibility testing of Aspergillus species against four commonly used antifungals.

机构信息

Department of Microbiology, Laboratory Medicine, Royal Brompton and Harefield NHS foundation trust, London.

Department of Microbiology, Kings College Hospital, London.

出版信息

Med Mycol. 2020 Nov 10;58(8):1085-1090. doi: 10.1093/mmy/myaa020.

DOI:10.1093/mmy/myaa020
PMID:32277834
Abstract

The aim of this study was to evaluate a colorimetric method, MIRONAUT-AM, for determining susceptibility testing of anidulafungin, amphotericin, voriconazole, and itraconazole by comparing the minimum inhibitory (effective) concentrations (MICs/MECs) obtained by this method to those generated by the reference Clinical Laboratory Standard Institute (CLSI) broth microdilution method. In sum, 78 clinical isolates of Aspergillus species, nine of them non-wild type (non-WT) with itraconazole MIC ranging from 2 mg/l to >16 mg/l, were tested against above antifungals. A. fumigatus ATCC 204305 was used as a reference strain, and test was performed in accordance with slightly modified yeast susceptibility testing instruction of the manufacture; conidia suspension inoculum and alamarBlue concentration were optimized. These same isolates were referred to Bristol Mycology reference laboratory and tested by CLSI method. The MICs and MECs generated by the two methods were compared using concordance analysis. MIRONAUT-AM showed significant concordance (P < .0001) with CLSI method, and overall agreement was high (≥90%). In addition, MIRONAUT-AM produced echinocandin MECs results within 18-24 hours incubation time and correctly detected all non-WT isolates except one isolate. This colorimetric method is very promising and appears to be a suitable alternative susceptibility testing method to labor intensive broth microdilution reference method for Aspergillus species.

摘要

本研究旨在评估比色法 MIRONAUT-AM 用于测定棘白菌素(anidulafungin)、两性霉素、伏立康唑和伊曲康唑的药敏试验,通过比较该方法获得的最小抑菌(有效)浓度(MIC/MEC)与参考临床实验室标准化协会(CLSI)肉汤微量稀释法生成的 MIC/MEC。本研究共检测了 78 株临床分离的曲霉菌属菌株,其中 9 株为伊曲康唑 MIC 值在 2mg/L 至>16mg/L 之间的非野生型(non-WT)菌株,对上述抗真菌药物进行了药敏试验。烟曲霉 ATCC 204305 被用作参考菌株,检测方法根据生产商稍作修改的酵母菌药敏检测说明进行,优化了孢子悬浮液接种物和 alamarBlue 浓度。这些相同的分离株被送到 Bristol 真菌学参考实验室,并用 CLSI 方法进行检测。使用一致性分析比较两种方法生成的 MIC 和 MEC。MIRONAUT-AM 与 CLSI 方法具有显著的一致性(P<0.0001),总符合率较高(≥90%)。此外,MIRONAUT-AM 在 18-24 小时孵育时间内产生棘白菌素 MEC 结果,并正确检测除一株以外的所有非野生型分离株。这种比色法非常有前途,似乎是一种替代劳动密集型肉汤微量稀释参考方法的合适的药敏试验方法,适用于曲霉菌属。

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