Group for Biomedical Research in Sepsis (Bio Sepsis), Hospital Clínico Universitario de Valladolid/IECSCYL, Valladolid, Spain.
Anesthesiology and Reanimation Service, Hospital Clínico Universitario de Valladolid, Valladolid, Spain.
Ann Surg. 2019 Mar;269(3):545-553. doi: 10.1097/SLA.0000000000002406.
To quantify immunological dysfunction in surgical patients with presence/absence of sepsis using a droplet digital polymerase chain reaction (ddPCR) transcriptomic analysis. The study also aims to evaluate this approach for improving identification of sepsis in these patients.
Immune dysregulation is a central event in sepsis. Quantification of the expression of immunological genes participating in the pathogenesis of sepsis could represent a new avenue to improve its diagnosis.
Expression of 6 neutrophil protease genes (MMP8, OLFM4, LCN2/NGAL, LTF, PRTN3, MPO) and also of 5 genes involved in the immunological synapse (HLA-DRA, CD40LG, CD3E, CD28, ICOS) was quantified in blood from 101 surgical patients with sepsis, 53 uninfected surgical patients, and 16 blood donors by using ddPCR. Areas under receiver operating characteristic curves (AUROC) and multivariate regression analysis were employed to test individual genes and gene ratios to identify sepsis, in comparison with procalcitonin.
Sepsis-induced overexpression of neutrophil protease genes and depressed expression of immunological synapse genes. MMP8/HLA-DRA, LCN2/HLA-DRA outperformed procalcitonin in differentiating between patients with sepsis and surgical controls in the AUROC analysis: LCN2/HLA-DRA: 0.90 (0.85-0.96), MMP8/HLA-DRA: 0.89 (0.84-0.95), procalcitonin: 0.80 (0.73-0.88) (AUROC, confidence interval 95%), and also in the multivariate analysis: LCN2/HLA-DRA: 8.57 (2.25-32.62); MMP8/HLA-DRA: 8.03 (2.10-30.76), procalcitonin: 4.20 (1.15-15.43) [odds ratio (confidence interval 95%)]. Gene expression levels of HLA-DRA were an independent marker of hospital mortality.
Quantifying the transcriptomic ratios MMP8/HLA-DRA, LCN2/HLA-DRA by ddPCR is a promising approach to improve sepsis diagnosis in surgical patients.
使用液滴数字聚合酶链反应（ddPCR）转录组分析量化存在/不存在脓毒症的外科患者的免疫功能障碍。该研究还旨在评估这种方法在提高这些患者脓毒症识别方面的作用。
免疫失调是脓毒症的核心事件。量化参与脓毒症发病机制的免疫基因的表达可能代表改善其诊断的新途径。
通过 ddPCR 定量检测 101 例脓毒症手术患者、53 例未感染手术患者和 16 名献血者血液中 6 种中性粒细胞蛋白酶基因（MMP8、OLFM4、LCN2/NGAL、LTF、PRTN3、MPO）和 5 种参与免疫突触的基因（HLA-DRA、CD40LG、CD3E、CD28、ICOS）的表达。使用受试者工作特征曲线（AUROC）和多变量回归分析测试单个基因和基因比值，以确定与降钙素相比脓毒症的诊断。
脓毒症诱导的中性粒细胞蛋白酶基因过度表达和免疫突触基因表达下调。在 AUROC 分析中，MMP8/HLA-DRA 和 LCN2/HLA-DRA 优于降钙素区分脓毒症患者和手术对照组：LCN2/HLA-DRA：0.90（0.85-0.96），MMP8/HLA-DRA：0.89（0.84-0.95），降钙素：0.80（0.73-0.88）（AUROC，95%置信区间），并且在多变量分析中也是如此：LCN2/HLA-DRA：8.57（2.25-32.62）；MMP8/HLA-DRA：8.03（2.10-30.76），降钙素：4.20（1.15-15.43）[比值比（95%置信区间）]。HLA-DRA 的基因表达水平是医院死亡率的独立标志物。
通过 ddPCR 定量 MMP8/HLA-DRA、LCN2/HLA-DRA 的转录组比值是改善外科患者脓毒症诊断的一种很有前途的方法。
