乳铁蛋白-DNA复合物检测评价甲型H1N1重症流感患者血浆中性粒细胞胞外诱捕网水平  

Lactoferrin-DNA complex detection to evaluate plasma neutrophil extracellular traps of patients with severe influenza A (H1N1)

作　　者：杨晓宇 张玥[2,3] 麦维利 李蕊 朱鏐娈[1,2,3] 韩俊燕 曾辉 Yang Xiaoyu;Zhang yue;Mai Weili;Li Rui;Zhu Liuluan;Han Junyan;Zeng Hui(Peking University Ditan Teaching Hospital,Beijing 100015,China;Institute of Infectious Diseases,Beijing Ditan Hospital,Capital Medical University,Beijing 100015,China;Beijing Key Laboratory of Emerging Infectious Diseases,Beijing 100015,China)

机构地区：[1]北京大学地坛医院教学医院,北京100015 [2]首都医科大学附属北京地坛医院传染病研究所,北京100015 [3]新发突发传染病研究北京市重点实验室,北京100015

出　　处：《中华实验和临床感染病杂志（电子版）》2019年第2期122-127,共6页Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)

基　　金：国家自然科学基金(No.81570372;No.81501692)

摘　　要：目的建立乳铁蛋白-DNA复合物的相对定量检测方法,评价甲型H1N1重型流感及其他病原体感染者血浆样本中性粒细胞胞外诱捕网(NETs)的含量。方法将DNA含量明确的血浆样本进行梯度稀释作为标准品,应用酶联免疫吸附法(ELISA)检测血浆中乳铁蛋白-DNA复合物含量。采用乳铁蛋白-DNAELISA方法比较19例甲型H1N1重症流感患者与19例献血者(对照组)血浆乳铁蛋白-DNA复合物含量差异。结果甲型H1N1重型重症流感患者血浆中乳铁蛋白-DNA复合物水平与游离DNA(cfDNA)和组蛋白-DNA复合物水平均呈正相关(r=0.6763、P<0.0001,r=0.7560、P<0.0001)。甲型H1N1重症流感患者血浆中乳铁蛋白-DNA复合物含量为[2.082(1.169,5.021)μg/ml],显著高于对照组[0.233(0.170,0.376)μg/ml],差异有统计学意义(P<0.0001),且乳铁蛋白-DNA复合物含量与甲型H1N1重症流感患者的急性生理与慢性健康评分(APACHEⅡ评分)呈正相关(r=0.7379、P=0.0005)。结论血浆乳铁蛋白-DNA复合物含量可用于评价甲型H1N1重症流感患者NETs水平。Objective To establish a relative quantitative assay for lactoferrin-DNA complex, and to assess the levels of neutrophil extracellular traps(NETs)in plasma samples from patients with severe influenza A(H1N1) and other pathogen infectious diseases. Methods Plasma samples with clear DNA content were serially diluted as a standard, and the level of lactoferrin-DNA complex in plasma was determined by Enzymelinked immunosorbent assay(ELISA). The lactoferrin-DNA ELISA method was used to compare the difference of plasma lactoferrin-DNA complex between 19 patients with severe influenza A(H1N1) and 19 blood donors(control group). Results Level of plasma lactoferrin-DNA complex of patients in severe influenza A(H1N1)was positively correlated with that of cfDNA(cell-free DNA) and histone-DNA complex, respectively(r =0.6763, P < 0.0001;r = 0.756, P < 0.0001). The plasma lactoferrin-DNA complex content in patients with severe influenza A(H1N1) was 2.082(1.169, 5.021) μg/ml, which was significantly higher than that of the controls [0.233(0.170, 0.376) μg/ml, P < 0.0001], and positively correlated with Acute Physiological and Chronic Health Evaluation Ⅱ score(APACHE Ⅱ score)(r = 0.7379, P = 0.0005). Conclusion Plasma lactoferrin-DNA complex level could be used to assess NETs level in patients with severe influenza A(H1N1).

关 键 词：甲型H1N1重型流感 乳铁蛋白-DNA复合物 中性粒细胞胞外诱捕网 酶联免疫吸附法 

分 类 号：R511.7[医药卫生—内科学]