High-Throughput Identification of Bacteria and Yeast by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry in Conventional Medical Microbiology Laboratories
High-Throughput Identification of Bacteria and Yeast by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry in Conventional Medical Microbiology Laboratories
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DOI：
10.1128/jcm.02071-09
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发表时间：
2010-03-01
影响因子：
9.4
通讯作者：
Kuijper, Ed J.
中科院分区：
文献类型：
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作者：
van Veen, S. Q.;Claas, E. C. J.;Kuijper, Ed J.
基质辅助激光解吸电离飞行时间质谱仪(MALDI-TOF MS)适合于高通量、低成本的快速诊断，可以被认为是传统微生物实验室常规生化和分子鉴定系统的替代。首先，我们使用327种临床分离株对MALDI-TOF MS进行了评估，这些分离株以前是从患者材料中培养出来的，并通过常规技术(VITEK-II、API和生化测试)进行了鉴定。通过16S基因的分子分析对差异进行分析。利用MALDI-TOF MS对327株菌株进行了属水平和种水平的鉴定，准确率为95.1%。其次，对980株临床分离的细菌和酵母进行了前瞻性验证研究。MALDI-TOF MS对物种鉴定的正确率(分别为92.2%和83.1%)明显优于常规生化系统(分别为92.2%和83.1%)，错误属鉴定的准确率(分别为0.1%和1.6%)明显低于常规生化系统。MALDI-TOF MS鉴定准确率为97.7%的肠杆菌科细菌、92%的非发酵革兰氏阴性菌、94.3%的葡萄球菌、84.8%的链球菌、84%的杂类(主要是嗜血杆菌、放线杆菌、心脏杆菌、艾肯氏杆菌和金黄色葡萄球菌)和85.2%的酵母菌。MALDI-TOF MS在葡萄球菌的种类鉴定和Hacek属细菌的属鉴定方面明显优于常规方法。MALDI-TOF MS的错误鉴定显然与MALDI-TOF MS数据库中合适的参考菌株缺乏足够的光谱有关。我们得出结论，MALDI-TOF MS可以方便地用于医学微生物学实验室中细菌(肺炎球菌和绿色链球菌除外)和酵母菌的常规鉴定。
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.