Markanović Manda, Makek Mateja Janković, Glodić Goran, Kuliš Tomislav, Mareković Ivana
Department of Clinical and Molecular Microbiology, University Hospital Centre Zagreb, Zagreb, 10 000, Croatia.
School of Medicine, University of Zagreb, Zagreb, 10 000, Croatia.
J Mass Spectrom. 2023 Apr;58(4):e4915. doi: 10.1002/jms.4915.
Major challenges in the identification of non-tuberculous mycobacteria (NTM) by MALDI-TOF MS include protein extraction protocol and updating of the NTM database. The aim of this study was to evaluate MALDI Biotyper Mycobacteria Library v6.0 (Bruker Daltonics GmbH, Bremen, Germany) for identification of clinical NTM isolates and its impact on clinical management. NTM isolates cultivated from clinical samples in 101 patients were identified simultaneously by PCR-reverse hybridization (Hain Lifescience GmbH, Nehren, Germany) as a routinely used reference molecular method and using MALDI Biotyper Microflex LT/SH after protein extraction. Each isolate was applied to eight spots, and mean scores were used in analysis. MALDI-TOF MS obtained correct identification to the species level for 95 (94.06%) NTM isolates. The majority of correctly identified isolates (92/95; 96.84%) were identified with high-confidence score of ≥1.80 and only 3.16% (3/95) with a score of <1.80. Mean value ± SD of RGM NTM isolates (2.127 ± 0.172) was statistically significant higher in comparison to SGM NTM isolates (2.027 ± 0.142) with a p value of 0.007. In comparison to PCR-reverse hybridization, discordant identification results by MALDI-TOF MS were found in six (6/101; 5.94%) NTM isolates for which clinical data were analyzed. We demonstrated a high confidence NTM identifications using Mycobacterium Library v 6.0 on routine clinical isolates. This is the first study that analyzed MALDI-TOF MS identification results of NTM isolates in the context of clinical data, and it showed that MALDI-TOF MS with its updated databases could help clarify the epidemiology, clinical characteristics, and course of infections caused by less frequent NTM species.
通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)鉴定非结核分枝杆菌(NTM)的主要挑战包括蛋白质提取方案和NTM数据库的更新。本研究的目的是评估MALDI Biotyper分枝杆菌文库v6.0(德国不来梅布鲁克道尔顿公司)用于鉴定临床NTM分离株及其对临床管理的影响。从101例患者临床样本中培养的NTM分离株,同时采用聚合酶链反应-反向杂交法(德国内伦海因生命科学公司)作为常规使用的参考分子方法进行鉴定,并在蛋白质提取后使用MALDI Biotyper Microflex LT/SH进行鉴定。每个分离株点样8个斑点,分析时使用平均得分。MALDI-TOF MS对95株(94.06%)NTM分离株实现了种水平的正确鉴定。大多数正确鉴定的分离株(92/95;96.84%)鉴定得分≥1.80,为高置信度,只有3.16%(3/95)得分<1.80。快速生长型NTM分离株的平均值±标准差(2.127±0.172)与缓慢生长型NTM分离株(2.027±0.142)相比具有统计学显著差异,p值为0.007。与聚合酶链反应-反向杂交法相比,MALDI-TOF MS在6株(6/101;5.94%)NTM分离株中发现了不一致的鉴定结果,并对这些分离株的临床数据进行了分析。我们证明了使用分枝杆菌文库v6.0对常规临床分离株进行NTM鉴定具有高置信度。这是第一项在临床数据背景下分析NTM分离株的MALDI-TOF MS鉴定结果的研究,结果表明,更新数据库后的MALDI-TOF MS有助于阐明由较少见的NTM菌种引起的感染的流行病学、临床特征和病程。