首页> 美国卫生研究院文献>Journal of Clinical Microbiology >Comparison of Sample Preparation Methods Instrumentation Platforms and Contemporary Commercial Databases for Identification of Clinically Relevant Mycobacteria by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry
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Comparison of Sample Preparation Methods Instrumentation Platforms and Contemporary Commercial Databases for Identification of Clinically Relevant Mycobacteria by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

机译:通过基质辅助激光解吸电离-飞行时间质谱鉴定临床相关分枝杆菌的样品制备方法仪器平台和当代商业数据库的比较

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摘要

When mycobacteria are recovered in clinical specimens, timely species-level identification is required to establish the clinical significance of the isolate and facilitate optimization of antimicrobial therapy. Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) has recently been reported to be a reliable and expedited method for identification of mycobacteria, although various specimen preparation techniques and databases for analysis are reported across studies. Here we compared two MALDI-TOF MS instrumentation platforms and three databases: Bruker Biotyper Real Time Classification 3.1 (Biotyper), Vitek MS Plus Saramis Premium (Saramis), and Vitek MS v3.0. We evaluated two sample preparation techniques and demonstrate that extraction methods are not interchangeable across different platforms or databases. Once testing parameters were established, a panel of 157 mycobacterial isolates (including 16 Mycobacterium tuberculosis isolates) was evaluated, demonstrating that with the appropriate specimen preparation, all three methods provide reliable identification for most species. Using a score cutoff value of ≥1.8, the Biotyper correctly identified 133 (84.7%) isolates with no misidentifications. Using a confidence value of ≥90%, Saramis correctly identified 134 (85.4%) isolates with one misidentification and Vitek MS v3.0 correctly identified 140 (89.2%) isolates with one misidentification. The levels of accuracy were not significantly different across the three platforms (P = 0.14). In addition, we show that Vitek MS v3.0 requires modestly fewer repeat analyses than the Biotyper and Saramis methods (P = 0.04), which may have implications for laboratory workflow.
机译:当临床标本中的分枝杆菌恢复后,需要及时进行种属水平鉴定,以建立分离株的临床意义并促进抗菌治疗的优化。最近,据报道基质辅助激光解吸电离-飞行时间质谱(MALDI-TOF MS)是一种可靠且快速的鉴定分枝杆菌的方法,尽管各种研究都报告了各种标本制备技术和分析数据库。在这里,我们比较了两个MALDI-TOF MS仪器平台和三个数据库:Bruker Biotyper实时分类3.1(Biotyper),Vitek MS Plus Saramis Premium(Saramis)和Vitek MS v3.0。我们评估了两种样品制备技术,并证明了提取方法在不同平台或数据库之间不可互换。建立测试参数后,将评估一组157个分枝杆菌菌株(包括16个结核分枝杆菌菌株),证明通过适当的样品制备,这三种方法均可为大多数物种提供可靠的鉴定。使用分数截止值≥1.8,Biotyper可以正确识别133个(84.7%)分离株,且没有错误鉴定。使用≥90%的置信度值,Saramis可以正确识别出134个(85.4%)带有一个错误识别的菌株,而Vitek MS v3.0可以正确识别出140个(89.2%)带有一个错误识别的菌株。在三个平台上,准确性水平没有显着差异(P = 0.14)。此外,我们显示Vitek MS v3.0与Biotyper和Saramis方法相比,所需重复分析的次数要少一些(P = 0.04),这可能对实验室工作流程有影响。

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