Assistance Publique-Hôpitaux de Paris, Laboratoire de Microbiologie, Hôpital Necker-Enfants Malades, and Université Paris 5 Descartes, Faculté de Medicine, Site Necker, 149 rue de Sèvres, 75015 Paris, France.
J Clin Microbiol. 2010 Dec;48(12):4481-6. doi: 10.1128/JCM.01397-10. Epub 2010 Oct 13.
Mycobacterial identification is based on several methods: conventional biochemical tests that require several weeks for accurate identification, and molecular tools that are now routinely used. However, these techniques are expensive and time-consuming. In this study, an alternative method was developed using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). This approach allows a characteristic mass spectral fingerprint to be obtained from whole inactivated mycobacterial cells. We engineered a strategy based on specific profiles in order to identify the most clinically relevant species of mycobacteria. To validate the mycobacterial database, a total of 311 strains belonging to 31 distinct species and 4 species complexes grown in Löwenstein-Jensen (LJ) and liquid (mycobacterium growth indicator tube [MGIT]) media were analyzed. No extraction step was required. Correct identifications were obtained for 97% of strains from LJ and 77% from MGIT media. No misidentification was noted. Our results, based on a very simple protocol, suggest that this system may represent a serious alternative for clinical laboratories to identify mycobacterial species.
传统的生化试验需要数周才能进行准确鉴定,而分子工具现在已常规使用。然而,这些技术既昂贵又耗时。在这项研究中,使用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)开发了一种替代方法。这种方法允许从整个失活分枝杆菌细胞中获得特征质谱指纹图谱。我们设计了一种基于特定图谱的策略,以鉴定最具临床相关性的分枝杆菌物种。为了验证分枝杆菌数据库,共分析了 311 株属于 31 个不同物种和 4 个种复合体的分枝杆菌,分别在 Löwenstein-Jensen(LJ)和液体(分枝杆菌生长指示剂管 [MGIT])培养基中生长。不需要提取步骤。从 LJ 中获得了 97%的菌株的正确鉴定,从 MGIT 培养基中获得了 77%的正确鉴定。没有错误鉴定。我们的结果基于一个非常简单的方案,表明该系统可能代表临床实验室鉴定分枝杆菌物种的一种可行替代方法。