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
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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DOI：
10.1128/jcm.00567-15
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发表时间：
2015-07-01
影响因子：
9.4
通讯作者：
Burnham, Carey-Ann D.
中科院分区：
文献类型：
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作者：
Wilen, Craig B.;McMullen, Allison R.;Burnham, Carey-Ann D.
当临床标本中发现分枝杆菌时，需要及时进行物种水平鉴定，以确定分离株的临床意义并促进优化抗菌治疗。尽管研究中报道了各种样本制备技术和分析数据库，但最近报道称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分枝杆菌鉴定方法。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 Real Time Classification 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，并证明提取方法在不同平台或数据库之间不可互换。确定测试参数后，将对 157 种分枝杆菌分离株（包括 16 种结核分枝杆菌分离株）进行评估，证明通过适当的标本制备，所有三种方法都能为大多数物种提供可靠的鉴定。使用 >= 1.8 的分数截止值，Biotyper 正确识别了 133 个分离株 (84.7%)，没有错误识别。使用 >= 90% 的置信度值，Saramis 正确识别出 134 个（85.4%）个带有 1 个错误识别的分离株，Vitek MS v3.0 正确识别出 140 个（89.2%）个带有 1 个错误识别的分离株。三个平台的准确度水平没有显着差异（P = 0.14）。此外，我们还发现 Vitek MS v3.0 所需的重复分析次数比 Biotyper 和 Saramis 方法略少 (P = 0.04)，这可能对实验室工作流程产生影响。
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.