Clinic for Avian, Reptile and Fish Medicine, Department for Farm Animals and Veterinary Public Health, University of Veterinary Medicine, Vienna, Austria.
VetOMICS Core Facility for Research/Proteomics and Metabolomics, University of Veterinary Medicine, Vienna, Austria.
J Med Microbiol. 2010 Mar;59(Pt 3):295-301. doi: 10.1099/jmm.0.016576-0. Epub 2009 Dec 3.
Rapid and reliable identification of Arcobacter and Helicobacter species, and their distinction from phenotypically similar Campylobacter species, has become increasingly important, since many of them are now recognized as human and/or animal pathogens. Matrix-associated laser desorption/ionization-time of flight (MALDI-TOF) MS has been shown to be a rapid and sensitive method for characterization of micro-organisms. In this study, we therefore established a reference database of selected Arcobacter, Helicobacter and Campylobacter species for MALDI-TOF MS identification. Besides the species with significance as food-borne pathogens - Arcobacter butzleri, Helicobacter pullorum, Campylobacter jejuni and Campylobacter coli - several other members of these genera were included in the reference library to determine the species specificity of the designed MALDI Biotyper reference database library. Strains that made up the reference database library were grown on Columbia agar, and yielded reproducible and unique mass spectra profiles, which were compared with the Bruker Biotyper database, version 2. The database was used to identify 144 clinical isolates using whole spectral profiles. Furthermore, reproducibility of MALDI-TOF MS results was evaluated with respect to age and/or storage of bacteria and different growth media. It was found that correct identification could be obtained even if the bacteria were stored at room temperature or at 4 degrees C up to 9 days before being tested. In addition, bacteria were correctly identified when grown on Campylosel agar; however, they were not when grown on modified charcoal cefoperazone deoxycholate agar. These results indicate that MALDI-TOF MS fingerprinting is a fast and reliable method for the identification of Arcobacter and Helicobacter species, and their distinction from phenotypically similar Campylobacter species, with applications in clinical diagnostics.
快速可靠地鉴定弯曲菌属和螺杆菌属物种，以及将其与表型相似的弯曲杆菌属物种区分开来，变得越来越重要，因为许多这些物种现在被认为是人类和/或动物病原体。基质辅助激光解吸/电离飞行时间（MALDI-TOF）MS 已被证明是一种快速而敏感的微生物鉴定方法。因此，在本研究中，我们建立了一个选定的弯曲菌属、螺杆菌属和弯曲杆菌属物种的 MALDI-TOF MS 鉴定参考数据库。除了作为食源性病原体具有重要意义的物种（弯曲菌属 butzleri、鸡螺杆菌、空肠弯曲菌和大肠弯曲菌）外，该参考文库还包括这些属的其他几个成员，以确定设计的 MALDI Biotyper 参考数据库文库的物种特异性。参考数据库库中的菌株在哥伦比亚琼脂上生长，产生可重复且独特的质谱谱图，这些谱图与 Bruker Biotyper 数据库版本 2 进行比较。该数据库用于使用全谱图鉴定 144 株临床分离株。此外，还评估了 MALDI-TOF MS 结果的重现性，包括细菌的年龄和/或储存以及不同的生长培养基。结果发现，即使将细菌在室温下或在 4°C 下储存长达 9 天，在进行测试之前，也可以获得正确的鉴定结果。此外，当在弯曲菌琼脂上生长时，细菌可以被正确鉴定；然而，当在改良的炭头孢哌酮去氧胆酸盐琼脂上生长时，它们则不能被正确鉴定。这些结果表明，MALDI-TOF MS 指纹图谱是一种快速可靠的鉴定弯曲菌属和螺杆菌属物种以及将其与表型相似的弯曲杆菌属物种区分开来的方法，可应用于临床诊断。
