Clínica El Rosario, Carrera 41 #62-5, Medellín, Colombia; Corporación para Investigaciones Biológicas (CIB), Cra. 72 A No. 78 B 141, Medellín, Colombia; Hospital general de Medellín "Luz Castro de Gutierrez" ESE, Carrera 48 # 32-102, Medellín, Colombia.
Laboratorio Médico de Referencia, Calle 63 No. 41-27, Medellín, Colombia.
J Glob Antimicrob Resist. 2018 Jun;13:254-260. doi: 10.1016/j.jgar.2017.11.010. Epub 2017 Nov 26.
Invasive candidiasis has a high impact on morbidity and mortality in hospitalised patients. Accurate and timely methods for identification of Candida spp. and determination of echinocandin susceptibility have become a priority for clinical microbiology laboratories.
This study was performed to compare matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS) identification with sequencing of the D1/D2 region of the rRNA gene complex 28 subunit in 147 Candida spp. isolates obtained from patients with candidaemia. Antimicrobial susceptibility testing was performed by broth microdilution (BMD) and Etest. Sequencing of the FKS1 and FKS2 genes was performed.
The most common species isolated were Candida albicans (40.8%), followed by Candida parapsilosis (23.1%) and Candida tropicalis (17.0%). Overall agreement between the results of identification by MALDI-TOF/MS and molecular identification was 99.3%. Anidulafungin and caspofungin susceptibility by the BMD method was 98.0% and 88.4%, respectively. Susceptibility to anidulafungin and caspofungin by Etest was 93.9% and 98.6%, respectively. Categorical agreement between Etest and BMD was 91.8% for anidulafungin and 89.8% for caspofungin, with lower agreements in C. parapsilosis for anidulafungin (76.5%) and C. glabrata for caspofungin (40.0%). No mutations related to resistance were found in the FKS genes, although 54 isolates presented synonymous polymorphisms in the hotspots sequenced.
MALDI-TOF/MS is a good alternative for routine identification of Candida spp. isolates. DNA sequencing of the FKS genes suggested that the isolates analysed were susceptible to echinocandins; alternatively, unknown resistance mechanisms or limitations related to antifungal susceptibility tests may explain the resistance found in a few isolates.
侵袭性念珠菌病对住院患者的发病率和死亡率有很大影响。准确、及时地鉴定念珠菌属和确定棘白菌素敏感性已成为临床微生物学实验室的优先事项。
本研究比较了基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF/MS)鉴定与 147 株念珠菌属分离株的核糖体 RNA 基因复合物 28 亚基 D1/D2 区测序。通过肉汤微量稀释法(BMD)和 Etest 进行抗微生物药敏试验。进行 FKS1 和 FKS2 基因测序。
分离的最常见物种是白色念珠菌(40.8%),其次是近平滑念珠菌(23.1%)和热带念珠菌(17.0%)。MALDI-TOF/MS 鉴定与分子鉴定结果的总符合率为 99.3%。BMD 法测定的阿尼达卢单抗和卡泊芬净敏感性分别为 98.0%和 88.4%。Etest 的阿尼达卢单抗和卡泊芬净敏感性分别为 93.9%和 98.6%。Etest 与 BMD 的类别一致性分别为阿尼达卢单抗 91.8%和卡泊芬净 89.8%,而阿尼达卢单抗在近平滑念珠菌中的一致性较低(76.5%),卡泊芬净在光滑念珠菌中的一致性较低(40.0%)。未发现 FKS 基因与耐药性相关的突变,尽管在测序的热点中有 54 株分离株存在同义多态性。
MALDI-TOF/MS 是念珠菌属分离株常规鉴定的良好选择。FKS 基因的 DNA 测序表明,分析的分离株对棘白菌素敏感;或者,未知的耐药机制或与抗真菌药敏试验相关的局限性可能解释了少数分离株的耐药性。