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[广泛使用的方法、新分子方法和基质辅助激光解吸电离飞行时间质谱在念珠菌菌株常规诊断中的成本、可靠性及结果报告时间比较]

[A comparison of the costs, reliability and time of result periods of widely used methods, new molecular methods and MALDI TOF-MS in the routine diagnosis of Candida strains].

作者信息

Kal Çakmaklıoğulları Elçin, Aşgın Nergis, Değerli Kenan

机构信息

Karabük University Faculty of Medicine, Department of Medical Microbiology, Karabük, Turkey.

Celal Bayar University Faculty of Medicine, Department of Medical Microbiology, Manisa, Turkey.

出版信息

Mikrobiyol Bul. 2019 Apr;53(2):204-212. doi: 10.5578/mb.67952.

Abstract

In recent years, the fast and accurate identification of the Candida species is of great importance as the response to antifungal treatment differs among species. Following the treatment of several immunosuppressive diseases, fungal infections can emerge. The aim of this study was to compare the accuracy, costs and time of result periods of the methods used in the identification of the most common human fungal infectious agent, Candida strains. From various clinical samples sent to the Microbiology Laboratory of Karabuk University Training and Research Hospital between July 2016-December 2017, a total of 91 yeast isolates cultivated in blood agar (Becton Dickinson, USA) and/or Sabouraud dextrose agar (SDA-Oxoid, UK), confirmed with colony morphology and microscopic appearance, identified as Candida species with a fully automated identification system (Phoenix™ Yeast ID Panel, Becton Dickinson Diagnostics, USA) were included in the study. All the samples were examined with sequence analysis using ITS1 forward 5'-TCC GTA GGT GAA CCT GCG G-3' and ITS4 reverse 5'-TCC TCC GCT TAT TGA TAT GC-3' primers (Iontek, Turkey) and the matrix-assisted laser desorption-ionisation time of flight mass spectrometry (MALDI TOF-MS) systems. Molecular sequence analysis was accepted as the gold standard method and the results were compared with those of the other methods MALDI TOF-MS and Phoenix™ Yeast ID Panel in respect of the accuracy of the identification of Candida strains. According to the results of the DNA sequence analysis of the 91 Candida isolates included in the study, 24 were identified as Candida albicans, 20 Candida tropicalis, 16 Candida parapsilosis, 13 Candida glabrata, seven Candida kefyr, six Candida krusei, two of each Candida dubliniensis, Candida guilliermondi and one Candida lusitaniae. Compared to the results of the DNA sequence analysis, the accurate identification of the fully automated Phoenix™ system and the MALDI TOF-MS system was found as 92.3% and 97.8%, respectively. In addition to accuracy, costs and time of result periods of the three methods were also compared. Disregarding the cost of the device in the 3 methods, when the comparison was made of the cost per test and the time to results after pure production in SDA agar, the MALDI TOF-MS system was determined to have the lowest costs and provided results in the shortest time. As some of the Candida strains have antifungal resistance, identification of the strains must be a priority in respect of starting early treatment. The MALDI TOF-MS system has high performance in accurate identification, low costs and the system provides the results within minutes, thereby allowing immediate decision to be made for the antifungal treatment to be started. Thus, the morbidity, mortality and cost rates will be reduced. In conclusion, as the MALDI TOF-MS is a rapid, reliable and low cost per test system, it can be considered suitable for routine use in laboratories.

摘要

近年来,由于不同念珠菌物种对抗真菌治疗的反应不同,快速准确地鉴定念珠菌物种至关重要。在几种免疫抑制性疾病的治疗之后,可能会出现真菌感染。本研究的目的是比较用于鉴定最常见的人类真菌病原体念珠菌菌株的方法在准确性、成本和出结果时间方面的差异。从2016年7月至2017年12月送往卡拉布克大学培训与研究医院微生物实验室的各种临床样本中,选取了总共91株在血琼脂(美国BD公司)和/或沙氏葡萄糖琼脂(英国Oxoid公司的SDA)上培养的酵母分离株,通过菌落形态和显微镜外观进行确认,并使用全自动鉴定系统(美国BD诊断公司的Phoenix™酵母鉴定板)鉴定为念珠菌物种,纳入本研究。所有样本均使用ITS1正向引物5'-TCC GTA GGT GAA CCT GCG G-3'和ITS4反向引物5'-TCC TCC GCT TAT TGA TAT GC-3'(土耳其Iontek公司)进行序列分析以及基质辅助激光解吸电离飞行时间质谱(MALDI TOF-MS)系统检测。分子序列分析被视为金标准方法,并将念珠菌菌株鉴定的准确性结果与其他方法(MALDI TOF-MS和Phoenix™酵母鉴定板)进行比较。根据本研究中纳入的91株念珠菌分离株的DNA序列分析结果,24株被鉴定为白色念珠菌,20株为热带念珠菌,16株为近平滑念珠菌,13株为光滑念珠菌,7株为凯菲念珠菌,6株为克柔念珠菌,都柏林念珠菌和季也蒙念珠菌各2株,葡萄牙念珠菌1株。与DNA序列分析结果相比,全自动Phoenix™系统和MALDI TOF-MS系统的准确鉴定率分别为92.3%和97.8%。除了准确性之外,还比较了这三种方法的成本和出结果时间。在不考虑三种方法中设备成本的情况下,当比较SDA琼脂纯培养后的每次检测成本和出结果时间时,MALDI TOF-MS系统被确定成本最低且出结果时间最短。由于一些念珠菌菌株具有抗真菌耐药性,在开始早期治疗方面,菌株鉴定必须是首要任务。MALDI TOF-MS系统在准确鉴定方面具有高性能、成本低,并且该系统在几分钟内就能给出结果,从而可以立即决定开始抗真菌治疗。因此,发病率、死亡率和成本率将会降低。总之,由于MALDI TOF-MS是一种快速、可靠且每次检测成本低的系统,它可被认为适合在实验室常规使用。

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