A simple method for rapid microbial identification from positive monomicrobial blood culture bottles through matrix-assisted laser desorption ionization time-of-flight mass spectrometry
A simple method for rapid microbial identification from positive monomicrobial blood culture bottles through matrix-assisted laser desorption ionization time-of-flight mass spectrometry
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DOI：
10.1016/j.jmii.2017.03.005
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发表时间：
2018-10-01
影响因子：
7.4
通讯作者：
Lu, Jang-Jih
中科院分区：
文献类型：
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作者：
Lin, Jung-Fu;Ge, Mao-Cheng;Lu, Jang-Jih
背景和目的：快速识别血液中的微生物对于败血症的治疗至关重要，因为败血症的严重程度很高，通过基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）从阳性血培养瓶中直接识别微生物可以缩短周转时间。因此，我们开发了一种使用 MALDI-TOF MS 从阳性血培养物中快速进行微生物鉴定的简单方法。方法：我们修改了先前开发的方法，提出了一种更快、更简单和更经济的方法，其中包括离心和溶血。具体来说，我们的方法包括在 600g 和 3000g 重力加速度 (g) 下进行两阶段离心，然后添加裂解缓冲液并再次进行 3000g 离心。结果：总共鉴定了 324 种单微生物细菌培养物。物种鉴定的成功率为81.8%，与其他复杂方法相当。革兰氏阴性需氧菌的鉴定成功率最高（85%），其次是革兰氏阳性需氧菌（78.2%）和厌氧菌（67%）。该方法所需时间不到10分钟，每次使用成本不到0.2美元，并且便于批量处理。结论：我们的结论是，该方法在微生物实验室的临床使用是可行的，并且可以作为治疗或进一步补充诊断测试的参考。版权所有(C) 2017，台湾微生物学会。由爱思唯尔台湾有限公司出版。
Background and purpose: Rapid identification of microbes in the bloodstream is crucial in managing septicemia because of its high disease severity, and direct identification from positive blood culture bottles through matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) can shorten the turnaround time. Therefore, we developed a simple method for rapid microbiological identification from positive blood cultures by using MALDI-TOF MS.Methods: We modified previously developed methods to propose a faster, simpler and more economical method, which includes centrifugation and hemolysis. Specifically, our method comprises two-stage centrifugation with gravitational acceleration (g) at 600g and 3000g, followed by the addition of a lysis buffer and another 3000g centrifugation.Results: In total, 324 monomicrobial bacterial cultures were identified. The success rate of species identification was 81.8%, which is comparable with other complex methods. The identification success rate was the highest for Gram-negative aerobes (85%), followed by Gram-positive aerobes (78.2%) and anaerobes (67%). The proposed method requires less than 10 min, costs less than US$0.2 per usage, and facilitates batch processing.Conclusion: We conclude that this method is feasible for clinical use in microbiology laboratories, and can serve as a reference for treatments or further complementary diagnostic testing. Copyright (C) 2017, Taiwan Society of Microbiology. Published by Elsevier Taiwan LLC.