Department of Biochemistry and Medical Biotechnology, Calcutta School of Tropical Medicine, Kolkata, West Bengal, India.
Department of Tropical Medicine, Calcutta School of Tropical Medicine, Kolkata, West Bengal, India.
Clin Exp Immunol. 2022 May 13;208(1):72-82. doi: 10.1093/cei/uxac018.
Dengue infection can result in simple dengue fever or life-threatening severe dengue. Early identification of severe patients is needed for proper disease management. Dengue infection was screened among 168 symptomatic patients by qRT-PCR, anti-dengue IgM, and IgG ELISA. Dengue patients were categorized according to WHO classification. Viral load and dengue serotypes were determined by qRT-PCR. Levels of acute-phase-proteins (SAP, SAA2; CRP and ApoA1), endothelial (Ang2, VEGF), coagulation (fibrinogen) markers were determined by sandwich ELISA/immunoturbidimetry/western-blotting. Hepatic (ALT, AST, ALP) and other blood biochemical parameters were studied by autoanalyzer and haematology cell counter. Statistical analysis and protein-protein-interaction network were performed by GraphPad-Prism and STRINGS database, respectively. Among 87 dengue patients, significantly higher levels of Ang2, VEGF, CRP, SAA2, ApoA1, AST, ALT, and AST/ALT ratio and low level of fibrinogen were detected in severe-dengue cases compared to dengue without warning-signs, with seven of them severely altered during febrile-phase. Higher fold-change of Ang2 and VEGF as well as decreased fibrinogen were observed among patients with haemorrhagic-manifestation, clinical-fluid accumulation and thrombocytopenia. Functional network analysis predicted Ang2, VEGF, and CRP to be functionally and physically connected and SAA2 and ApoA1 to be functioning together. Correlation analyses also validated this connectivity by a strong positive correlation between Ang2, VEGF, and CRP. PCA analysis followed by hierarchical clustering heatmap analysis segregated severe-dengue patients from the rest, with VEGF, Ang2, ApoA1, AST, and ALT clearly distinguishing the severe-dengue group. Thus, serum levels of VEGF, Ang2, ApoA1, AST, and ALT might act as potential biomarkers for predicting dengue severity during the early stage.
登革热感染可导致单纯登革热或危及生命的严重登革热。需要早期识别重症患者，以便进行适当的疾病管理。通过 qRT-PCR、抗登革热 IgM 和 IgG ELISA 对 168 例有症状患者进行登革热筛查。根据世界卫生组织分类对登革热患者进行分类。通过 qRT-PCR 确定病毒载量和登革热血清型。通过夹心 ELISA/免疫比浊法/免疫印迹法测定急性期蛋白 (SAP、SAA2；CRP 和 ApoA1)、内皮 (Ang2、VEGF)、凝血 (纤维蛋白原) 标志物。通过自动分析仪和血细胞计数器研究肝 (ALT、AST、ALP) 和其他血液生化参数。通过 GraphPad-Prism 进行统计分析和蛋白质-蛋白质相互作用网络分析，通过 STRINGS 数据库进行分析。在 87 例登革热患者中，与无预警信号的登革热患者相比，严重登革热患者的 Ang2、VEGF、CRP、SAA2、ApoA1、AST、ALT 和 AST/ALT 比值明显升高，且在发热期有 7 项指标严重改变。在有出血表现、临床液体积聚和血小板减少的患者中，Ang2 和 VEGF 的倍数变化较高，纤维蛋白原降低。功能网络分析预测 Ang2、VEGF 和 CRP 在功能和物理上相互连接，SAA2 和 ApoA1 一起发挥作用。相关性分析也通过 Ang2、VEGF 和 CRP 之间的强正相关验证了这种连接。PCA 分析后分层聚类热图分析将重症登革热患者与其他患者分开，VEGF、Ang2、ApoA1、AST 和 ALT 清楚地区分重症登革热组。因此，血清 VEGF、Ang2、ApoA1、AST 和 ALT 水平可能是早期预测登革热严重程度的潜在生物标志物。
