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通过基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)检测念珠菌属中的三唑耐药性。

Detection of triazole resistance among Candida species by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS).

作者信息

Saracli Mehmet A, Fothergill Annette W, Sutton Deanna A, Wiederhold Nathan P

机构信息

Gülhane Military Medical Academy, Ankara, Turkey

University of Texas Health Science Center at San Antonio, San Antonio, Texas.

出版信息

Med Mycol. 2015 Sep;53(7):736-42. doi: 10.1093/mmy/myv046. Epub 2015 Jul 10.

Abstract

MALDI-TOF MS can rapidly identify microorganisms to the species level and may be able to detect antimicrobial resistance. We evaluated the ability of this technology to detect triazole resistance in Candida species.35 C. albicans, 35 C. glabrata, and 37 C. tropicalis strains were exposed to fluconazole, voriconazole, or posaconazole at two different concentrations plus a drug-free control: a midrange concentration (CLSI clinical breakpoint or epidemiologic cut-off value), and a high concentration (fluconazole 64 μg/ml, voriconazole & posaconazole 16 μg/ml). The MALDI-TOF MS spectra at these concentrations were used to create the individual composite correlation index (CCI) matrices for each isolate. When the CCI of the midrange/highest concentration was lower than that of the midrange/null concentration, the strain was classified as resistant. These results were then compared to the classifications for susceptible or resistant obtained by measuring the MICs according to the CLSI M27-A3 antifungal susceptibility testing (AFST) method.The MALDI-TOF MS assay was able to classify triazole susceptibility against all strains. Overall, essential agreement between MALDI-TOF MS and AFST varied between 54% and 97%, and was highest for posaconazole against C. glabrata. The reproducibility of the MALDI-TOF MS assay varied between 54.3 and 82.9% and was best for fluconazole against C. albicans and posaconazole against C. glabrata. Reproducibility was also higher for C. glabrata isolates compared to C. albicans and C. tropicalis.These results demonstrate that MALDI-TOF MS may be used to simultaneously determine the Candida species and classification as susceptible or resistant to triazole antifungals. Further studies are needed to refine the methodology and improve the reproducibility of this assay.

摘要

基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)能够快速将微生物鉴定到种水平,并且或许能够检测抗菌药物耐药性。我们评估了该技术检测念珠菌属中三唑类耐药性的能力。35株白色念珠菌、35株光滑念珠菌和37株热带念珠菌菌株分别暴露于两种不同浓度的氟康唑、伏立康唑或泊沙康唑以及无药对照中:一个中等浓度(CLSI临床折点或流行病学截断值)和一个高浓度(氟康唑64μg/ml,伏立康唑和泊沙康唑16μg/ml)。这些浓度下的MALDI-TOF MS谱图用于为每个分离株创建单独的复合相关指数(CCI)矩阵。当中等浓度/最高浓度的CCI低于中等浓度/无药对照浓度的CCI时,该菌株被分类为耐药。然后将这些结果与根据CLSI M27-A3抗真菌药敏试验(AFST)方法测量最低抑菌浓度(MIC)获得的敏感或耐药分类进行比较。MALDI-TOF MS检测能够对所有菌株的三唑类药敏进行分类。总体而言,MALDI-TOF MS与AFST之间的基本一致性在54%至97%之间变化,泊沙康唑对光滑念珠菌的一致性最高。MALDI-TOF MS检测的重复性在54.3%至82.9%之间变化,氟康唑对白念珠菌和泊沙康唑对光滑念珠菌的重复性最佳。与白色念珠菌和热带念珠菌相比,光滑念珠菌分离株的重复性也更高。这些结果表明,MALDI-TOF MS可用于同时确定念珠菌属种类以及对三唑类抗真菌药物敏感或耐药的分类。需要进一步研究来完善该方法并提高该检测的重复性。

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