目的:本研究的主要目的是系统地筛选和鉴定败血症患者血浆中的差异表达蛋白(DEP)。这项工作采用了数据独立采集(DIA)和酶联免疫吸附测定(ELISA)方法。总体目标是提供可获得的和精确的血清生物标志物,有助于脓毒症的诊断识别。
方法:该研究包括2019年1月至2020年12月期间在西南医科大学附属医院收治的53例脓毒症患者,以及由16例无脓毒症病理的个体组成的对照队列。随后,包括来自对照组的10名随机选择的受试者和来自脓毒症组的22名受试者的子集经历经由DIA的定量蛋白质组学分析。获得的数据经过基因本体论(GO)和京都基因百科全书(KEGG)分析,促进蛋白质-蛋白质相互作用(PPI)网络的构建,以识别潜在的标记。然后通过ELISA完成核心蛋白的验证。随后的正常组和脓毒症组之间的比较分析,通过构建受试者工作特性(ROC)曲线来评估诊断效能。
结果:通过生物信息学方法共鉴定出187个DEP。检查显示它们主要参与生物过程,如伤口愈合,凝血,和血液凝固。功能通路分析进一步阐明了它们在补体通路和疟疾中的参与。抵抗素作为候选血浆生物标志物出现,随后通过ELISA验证。值得注意的是,与正常对照组相比,脓毒症患者血清中的蛋白质水平显着升高。ROC曲线分析强调了这些生物标志物对脓毒症的稳健诊断能力。
结论:数据独立采集(DIA)和酶联免疫吸附测定(ELISA)显示败血症患者的抵抗素水平升高,提示诊断潜力,保证进一步的研究。
OBJECTIVE: The primary objective of this investigation is to systematically screen and identify differentially expressed proteins (DEPs) within the plasma of individuals afflicted with sepsis. This endeavor employs both Data-Independent Acquisition (
DIA) and enzyme-linked immunosorbent assay (ELISA) methodologies. The overarching goal is to furnish accessible and precise serum biomarkers conducive to the diagnostic discernment of sepsis.
METHODS: The study encompasses 53 sepsis patients admitted to the Affiliated Hospital of Southwest Medical University between January 2019 and December 2020, alongside a control cohort consisting of 16 individuals devoid of sepsis pathology. Subsequently, a subset comprising 10 randomly selected subjects from the control group and 22 from the sepsis group undergoes quantitative proteomic analysis via
DIA. The acquired data undergoes Gene Ontology (GO) and Kyoto Encyclopedia of Genes (KEGG) analyses, facilitating the construction of a Protein-Protein Interaction (PPI) network to discern potential markers. Validation of core proteins is then accomplished through ELISA. Comparative analysis between the normal and sepsis groups ensues, characterized by Receiver Operating Characteristic (ROC) curve construction to evaluate diagnostic efficacy.
RESULTS: A total of 187 DEPs were identified through bioinformatic methodologies. Examination reveals their predominant involvement in biological processes such as wound healing, coagulation, and blood coagulation. Functional pathway analysis further elucidates their engagement in the complement pathway and malaria. Resistin emerges as a candidate plasma biomarker, subsequently validated through ELISA. Notably, the protein exhibits significantly elevated levels in the serum of sepsis patients compared to the normal control group. ROC curve analysis underscores the robust diagnostic capacity of these biomarkers for sepsis.
CONCLUSIONS: Data-Independent Acquisition (
DIA) and Enzyme-Linked Immunosorbent Assay (ELISA) show increased Resistin levels in sepsis patients, suggesting diagnostic potential, warranting further research.