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目的定量重症肝炎患者血浆DNA的水平,与ALT比较,评估鉴别重症肝炎的临床应用价值。方法收集30例重症肝炎患者外周血标本6ml,以急性肝炎(20例)、慢性乙肝(90例)、肝硬化(45例)和健康体检者(100例)作为对照。用磁珠法提取血浆DNA,双重实时荧光定量PCR测定血浆DNA。结果乙型肝炎患者的血浆DNA水平(中位数)显著高于健康对照(104.2ng/mlvs.23.4ng/ml,P=0.0000)。血浆DNA水平在重症肝炎患者和急性肝炎、慢性乙肝及肝硬化患者之间都具有显著性差异(P=0.0018、0.0000和0.0000)。乙型肝炎患者的血清ALT水平(中位数)显著高于健康对照(107.5U/Lvs.24.1U/L,P=0.0000)。重症肝炎患者的血清ALT水平与急性肝炎之间有显著差异(P=0.0024),但与慢性乙肝和肝硬化之间没有差异(P=0.0600和1.0000)。ROC曲线分析显示,在鉴别重症肝炎和肝硬化以及慢性乙肝时,血浆DNA的鉴别能力显著优于血清ALT(AUC,0.95vs.0.51,P=0.0000;0.86vs.0.34,P=0.0000)。结论用双重荧光定量PCR检测血浆DNA水平,可以作为一个用于准确鉴别重症肝炎有价值的生物学标志。
Objective To quantify the level of plasma DNA in patients with severe hepatitis and compare with ALT to evaluate the clinical value of identifying severe hepatitis. Methods Peripheral blood samples from 30 patients with severe hepatitis were collected. Sixteen acute hepatitis (20 cases), chronic hepatitis B (90 cases), cirrhosis (45 cases) and healthy subjects (100 cases) were collected as control. Plasma DNA was extracted by magnetic beads method and plasma DNA was detected by double real-time fluorescence quantitative PCR. Results Plasma DNA levels (median) in patients with hepatitis B were significantly higher than those in healthy controls (104.2 ng / ml vs 23.4 ng / ml, P = 0.0000). Plasma DNA levels were significantly different between patients with severe hepatitis and those with acute hepatitis, chronic hepatitis B and cirrhosis (P = 0.0018, 0.0000 and 0.0000). Serum ALT levels (median) in patients with hepatitis B were significantly higher than those in healthy controls (107.5 U / L vs 24.1 U / L, P = 0.0000). There was a significant difference between serum ALT levels and acute hepatitis in patients with severe hepatitis (P = 0.0024), but not between chronic hepatitis B and cirrhosis (P = 0.0600 and 1.0000). ROC curve analysis showed that the discrimination ability of plasma DNA was significantly better than that of serum ALT (AUC, 0.95vs.0.51, P = 0.0000; 0.86vs.0.34, P = 0.0000) in the identification of severe hepatitis and cirrhosis and chronic hepatitis B. Conclusions Dual-fluorescence quantitative PCR for detecting plasma DNA levels may serve as a valuable biomarker for the accurate identification of severe hepatitis.