【摘要】 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。更多还原

【Abstract】 Objectives To evaluate accurately hepatocyte injury degree of severe hepatitis patients by quantifying plasma DNA of severe hepatitis patients and study its clinical application in diagnosis of severe hepatitis comparing with ALT.Methods Six milliliters of peripheral blood samples were collected from 185 patients with hepatitis B which are divided into four groups, severe hepatitis with 30 cases, acute hepatitis with 20 cases, chronic B hepatitis with 90 cases, and liver cirrhosis with 45 cases. 100 healthy controls were enrolled. Circulating DNA was extracted from plasma by the BILATEST virus DNA/RNA extraction kit and quantified with a novel duplex real-time PCR assay, respectively.Results Plasma DNA levels of hepatitis B patients were significantly higher than those of healthy controls (104.2 ng/ml vs. 23.4 ng/ml (median),P=0.0000).A significant difference of plasma DNA concentration was found between severe hepatitis and acute hepatitis (P=0.0018), and chronic B hepatitis (P=0.0000), and liver cirrhosis (P=0.0000).The median value of serum ALT of hepatitis B patients was 107.5 U/L, much higher than that of the healthy controls (24.1 U/L,P=0.0000).The levels of serum ALT were significantly different between severe hepatitis and acute hepatitis (P=0.0024), while there was no remarkable difference between severe hepatitis and chronic B hepatitis (P=0.0600), liver cirrhosis (P=1.0000). Moreover, for distinguishing severe hepatitis from liver cirrhosis and chronic B hepatitis, the plasma DNA assay was notably superior to ALT by the analysis of receive operating characteristic (ROC) curves (AUC, 0.95 vs. 0.51,P=0.0000; 0.86 vs. 0.34,P=0.0000).Conclusion The results by measuring plasma DNA of hepatitis B patients with the novel duplex real-time quantitative PCR showed that plasma DNA may be considered as a robust predictive marker for accurately evaluating hepatocyte injury degree of severe hepatitis patients.更多还原