Rapid Inactivation of Mycobacterium and Nocardia Species before Identification Using Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry
Rapid Inactivation of Mycobacterium and Nocardia Species before Identification Using Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry
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DOI：
10.1128/jcm.01728-14
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发表时间：
2014-10-01
影响因子：
9.4
通讯作者：
Deol, Parampal
中科院分区：
文献类型：
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作者：
Dunne, W. Michael, Jr.;Doing, Kirk;Deol, Parampal
在生物控制设施外鉴定分枝杆菌需要首先使生物体处于不活跃状态。为了建立一种安全、有效、快速的灭活程序，该灭活程序与基质辅助激光解吸电离飞行时间质谱仪(MALDI-TOF MS)鉴定分枝杆菌和诺卡氏菌相兼容，对机械破坏前后暴露于70%乙醇(Etoh)的情况进行了评估。在70%乙醇中打珠5分钟，然后在室温下孵育10分钟的组合被发现具有快速杀菌作用，并且与直接在固体介质上生长获得的光谱相比，提供了高质量的光谱。培养物的年龄、冷藏或冷冻的裂解物的稳定性以及冻融循环不会对光谱的质量或所获得的鉴定产生不利影响。
The identification of mycobacteria outside biocontainment facilities requires that the organisms first be rendered inactive. Exposure to 70% ethanol (EtOH) either before or after mechanical disruption was evaluated in order to establish a safe, effective, and rapid inactivation protocol that is compatible with identification of Mycobacterium and Nocardia species using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). A combination of 5 min of bead beating in 70% EtOH followed by a 10-min room temperature incubation period was found to be rapidly bactericidal and provided high-quality spectra compared to spectra obtained directly from growth on solid media. The age of the culture, the stability of the refrigerated or frozen lysates, and freeze-thaw cycles did not adversely impact the quality of the spectra or the identification obtained.