Mayo Clinic Graduate School of Biomedical Sciences, Department of Immunology, Mayo Clinic, Rochester, Minnesota, USA.
Division of Clinical Microbiology, Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota, USA.
Proteomics Clin Appl. 2023 Sep;17(5):e2200071. doi: 10.1002/prca.202200071. Epub 2023 Mar 27.
This pilot study aimed to use proteomic profiling of sonicate fluid samples to compare host response during Staphylococcus aureus-associated periprosthetic joint infection (PJI) and non-infected arthroplasty failure (NIAF) and identify potential novel biomarkers differentiating the two.
In this pilot study, eight sonicate fluid samples (four from NIAF and four from S. aureus PJI) were studied. Samples were reduced, alkylated, and trypsinized overnight, followed by analysis using liquid chromatography-tandem mass spectrometry (LC-MS/MS) on a high-resolution Orbitrap Eclipse mass spectrometer. MaxQuant software suite was used for protein identification, filtering, and label-free quantitation.
Principal component analysis of the identified proteins clearly separated S. aureus PJI and NIAF samples. Overall, 810 proteins were identified based on their detection in at least three out of four samples from each group; 35 statistically significant differentially abundant proteins (DAPs) were found (two-sample t-test p-values ≤0.05 and log fold-change values ≥2 or ≤-2). Gene ontology pathway analysis found that microbial defense responses, specifically those related to neutrophil activation, to be increased in S. aureus PJI compared to NIAF samples.
Proteomic profiling of sonicate fluid using LC-MS/MS differentiated S. aureus PJI and NIAF in this pilot study. Further work is needed using a larger sample size and including non-S. aureus PJI and a diversty of NIAF-types.
本初步研究旨在利用超声液样本的蛋白质组学分析来比较金黄色葡萄球菌相关性假体周围关节感染(PJI)和非感染性关节置换失败(NIAF)期间的宿主反应,并确定潜在的新型生物标志物来区分两者。
在本初步研究中,研究了 8 个超声液样本(4 个来自 NIAF,4 个来自金黄色葡萄球菌 PJI)。样本经过还原、烷基化和过夜胰蛋白酶消化,然后在高分辨率 Orbitrap Eclipse 质谱仪上进行液相色谱-串联质谱(LC-MS/MS)分析。MaxQuant 软件套件用于蛋白质鉴定、过滤和无标记定量。
基于鉴定出的蛋白质的主成分分析清楚地将金黄色葡萄球菌 PJI 和 NIAF 样本分开。总体而言,基于每组至少有 4 个样本中的 3 个样本检测到的情况,鉴定出了 810 种蛋白质;发现了 35 种具有统计学意义的差异丰度蛋白(DAPs)(双样本 t 检验 p 值≤0.05,对数倍数变化值≥2 或≤-2)。基因本体论途径分析发现,与 NIAF 样本相比,金黄色葡萄球菌 PJI 中微生物防御反应增加,特别是与中性粒细胞激活相关的反应。
本初步研究中,使用 LC-MS/MS 对超声液进行蛋白质组学分析可区分金黄色葡萄球菌 PJI 和 NIAF。需要使用更大的样本量并包括非金黄色葡萄球菌 PJI 和多种 NIAF 类型来进一步开展工作。