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High-Throughput Identification of Bacteria and Yeast by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry in Conventional Medical Microbiology Laboratories

机译:传统医学微生物实验室中基质辅助激光解吸电离-飞行时间质谱法高通量鉴定细菌和酵母

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摘要

Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is suitable for high-throughput and rapid diagnostics at low costs and can be considered an alternative for conventional biochemical and molecular identification systems in a conventional microbiological laboratory. First, we evaluated MALDI-TOF MS using 327 clinical isolates previously cultured from patient materials and identified by conventional techniques (Vitek-II, API, and biochemical tests). Discrepancies were analyzed by molecular analysis of the 16S genes. Of 327 isolates, 95.1% were identified correctly to genus level, and 85.6% were identified to species level by MALDI-TOF MS. Second, we performed a prospective validation study, including 980 clinical isolates of bacteria and yeasts. Overall performance of MALDI-TOF MS was significantly better than conventional biochemical systems for correct species identification (92.2% and 83.1%, respectively) and produced fewer incorrect genus identifications (0.1% and 1.6%, respectively). Correct species identification by MALDI-TOF MS was observed in 97.7% of Enterobacteriaceae, 92% of nonfermentative Gram-negative bacteria, 94.3% of staphylococci, 84.8% of streptococci, 84% of a miscellaneous group (mainly Haemophilus, Actinobacillus, Cardiobacterium, Eikenella, and Kingella [HACEK]), and 85.2% of yeasts. MALDI-TOF MS had significantly better performance than conventional methods for species identification of staphylococci and genus identification of bacteria belonging to HACEK group. Misidentifications by MALDI-TOF MS were clearly associated with an absence of sufficient spectra from suitable reference strains in the MALDI-TOF MS database. We conclude that MALDI-TOF MS can be implemented easily for routine identification of bacteria (except for pneumococci and viridans streptococci) and yeasts in a medical microbiological laboratory.
机译:基质辅助激光解吸电离飞行时间质谱仪(MALDI-TOF MS)适用于低成本的高通量和快速诊断,可被视为常规微生物实验室中常规生化和分子鉴定系统的替代方案。首先,我们使用先前从患者材料中培养并通过常规技术(Vitek-II,API和生化测试)鉴定的327种临床分离株评估MALDI-TOF MS。通过对16S基因进行分子分析来分析差异。在327株分离物中,通过MALDI-TOF MS正确鉴定出95.1%的属水平,并且鉴定出85.6%的物种水平。其次,我们进行了一项前瞻性验证研究,包括980种细菌和酵母菌的临床分离株。 MALDI-TOF MS的总体性能明显优于传统生化系统,可进行正确的物种识别(分别为92.2%和83.1%),并且产生的错误属错误识别更少(分别为0.1%和1.6%)。通过MALDI-TOF MS对正确的物种进行鉴定,在97.7%的肠杆菌科细菌,92%的非发酵革兰氏阴性细菌,94.3%的葡萄球菌,84.8%的链球菌,84%的其他组(主要是嗜血杆菌,放线杆菌,心杆菌,艾肯氏菌)中观察到和Kingella [HACEK]),以及85.2%的酵母。 MALDI-TOF MS具有比常规方法更好的性能,可用于葡萄球菌的种类鉴定和HACEK组细菌的属鉴定。 MALDI-TOF MS的错误识别显然与MALDI-TOF MS数据库中合适的参考菌株缺乏足够的光谱有关。我们得出的结论是,在医学微生物实验室中,可以轻松实施MALDI-TOF MS常规检测细菌(除肺炎球菌和绿藻链球菌外)和酵母菌的常规方法。

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