Institute for Medical Microbiology, University Medical Center Göttingen, Göttingen, Germany.
Department of Microbiology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
Front Cell Infect Microbiol. 2021 Apr 19;11:634382. doi: 10.3389/fcimb.2021.634382. eCollection 2021.
Yeasts of the / species complexes are human pathogens mostly in immune compromised individuals, and can cause infections from dermal lesions to fungal meningitis. Differences in virulence and antifungal drug susceptibility of species in these complexes indicate the value of full differentiation to species level in diagnostic procedures. MALDI-TOF MS has been reported to sufficiently discriminate these species. Here, we sought to re-evaluate sample pre-processing procedures and create a set of publicly available references for use with the MALDI Biotyper system. Peak content using four different pre-processing protocols was assessed, and database entries for 13 reference strains created. These were evaluated against a collection of 153 clinical isolates, typed by conventional means. The use of decapsulating protocols or mechanical disruption did not sufficiently increase the information content to justify the extra hands-on-time. Using the set of 13 reference entries created with the standard formic acid extraction, we were able to correctly classify 143/153 (93.5%) of our test isolates. The majority of the remaining ten isolates still gave correct top matches; only two isolates did not give reproducible identifications. This indicates that the log score cut-off can be lowered also in this context. Ease to identify cryptococcal isolates to the species level is improved by the workflow evaluated here. The database references are freely available from https://github.com/oliverbader/BioTyper-libraries for incorporation into local diagnostic systems.
酵母/种复合物中的物种大多是免疫功能低下个体的人体病原体,可引起从皮肤损伤到真菌性脑膜炎的感染。这些复合物中种间的毒力和抗真菌药物敏感性差异表明,在诊断程序中充分区分到种水平具有重要价值。MALDI-TOF MS 已被报道足以区分这些种。在这里,我们试图重新评估样本预处理程序,并为 MALDI Biotyper 系统创建一组公开可用的参考资料。评估了使用四种不同预处理方案的峰含量,并为 13 个参考菌株创建了数据库条目。这些参考菌株与 153 株临床分离株进行了评估,这些分离株通过常规方法进行了分型。使用去包膜方案或机械破坏并不能充分增加信息量,因此不值得花费额外的时间。使用使用标准甲酸提取创建的 13 个参考条目集,我们能够正确分类 143/153(93.5%)的测试分离株。其余十个分离株中的大多数仍然给出了正确的顶级匹配;只有两个分离株没有给出可重复的鉴定。这表明在此背景下也可以降低对数得分截止值。通过这里评估的工作流程,易于将隐球菌分离株鉴定到种水平。数据库参考资料可从 https://github.com/oliverbader/BioTyper-libraries 免费获得,可用于整合到本地诊断系统中。