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基质辅助激光解吸电离飞行时间质谱技术(MALDI-TOF MS)对 BacT/Alert 血培养系统液体增菌后的标本进行快速直接检测,以诊断关节感染。

Rapid direct detection of pathogens for diagnosis of joint infections by MALDI-TOF MS after liquid enrichment in the BacT/Alert blood culture system.

机构信息

Institute of Medical Microbiology, University Medical Center Göttingen, Göttingen, Germany.

Department for Trauma Surgery, Orthopedics and Plastic Surgery, University Medical Center Göttingen, Göttingen, Germany.

出版信息

PLoS One. 2020 Dec 11;15(12):e0243790. doi: 10.1371/journal.pone.0243790. eCollection 2020.

Abstract

Pathogen identification is a critical step during diagnosis of infectious diseases. Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight mass spectrometry (MALDI-TOF-MS) has become the gold standard for identification of microorganisms cultured on solid media in microbiology laboratories. Direct identification of microbes from liquid specimen, circumventing the need for the additional overnight cultivation step, has been successfully established for blood culture, urine and liquor. Here, we evaluate the ability of MALDI-TOF MS for direct identification of pathogens in synovial fluid after liquid enrichment in BacT/Alert blood culture bottles. Influence of synovial specimen quality on direct species identification with the MALDI BioTyper/Sepsityper was tested with samples inoculated from pretested native synovia with concomitant inoculation of blood or pus, or highly viscous fluid. Here, we achieved >90% concordance with culture on solid medium, and only mixed-species samples posed significant problems. Performance in routine diagnostics was tested prospectively on bottles inoculated by treating physicians on ward. There, we achieved >70% concordance with culture on solid media. The major contributors to test failure were the absence of a measurable mass signal and mixed-specimen samples. The Sepsityper workflow worked well on samples derived from BacT/Alert blood culture bottles inoculated with synovial fluid, giving concordant results to identification from solid media. Host remnant material in the inoculum, such as blood or pus, had no detrimental effect on identification score values of the BioTyper system after processing with the Sepsityper workflow, and neither had the initial viscosity of the synovial sample.

摘要

病原体鉴定是传染病诊断过程中的关键步骤。基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)已成为微生物实验室中固体培养基上培养的微生物鉴定的金标准。直接从液体标本中鉴定微生物,避免了额外的过夜培养步骤,已经在血培养、尿液和体液中得到成功建立。在这里,我们评估了 MALDI-TOF MS 在 BacT/Alert 血培养瓶中液体富集后直接鉴定关节液中病原体的能力。通过对预先测试的天然关节液进行接种,并同时接种血液或脓液或高粘性液体,测试了 MALDI BioTyper/Sepsityper 对直接物种鉴定的关节液标本质量的影响。在门诊医生接种的瓶中进行前瞻性常规诊断性能测试。在那里,我们与固体培养基上的培养物达到了 >70%的一致性。只有混合物种样本造成了显著问题。Sepsityper 工作流程在接种了关节液的 BacT/Alert 血培养瓶中进行的样本上表现良好,与固体培养基上的鉴定结果一致。接种物中宿主残余物,如血液或脓液,在经过 Sepsityper 工作流程处理后,对 BioTyper 系统的鉴定评分值没有不利影响,初始关节液的粘度也没有影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/59f4/7732097/38efa3fef834/pone.0243790.g001.jpg

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