急性胰腺炎患者血浆脱氧核糖核酸定量分析及其临床意义

Quantitative analysis of circulating DNA in plasma of patients with acute pancreatitis and its potentialclinical value

目的 定量检测急性胰腺炎（AP）患者外周血血浆DNA并初步探讨其临床应用价值.方法 收集40例轻症急性胰腺炎（MAP）、20例重症急性胰腺炎（SAP）及50名健康志愿者的外周血血浆样本,采用微量基因组DNA提取试剂盒提取血浆DNA,以实时荧光定量PCR方法检测各组血浆DNA含量,比较分析血浆DNA水平与AP的临床相关性,并绘制受试者工作特征（ROC）曲线.结果 SAP组、MAP组和健康对照组血浆DNA含量的中位数分别为24.84、7.60和5.23ng/ml.60例AP患者血浆DNA含量明显高于健康对照组（P=0.006）,SAP组明显高于MAP组及健康对照组（P值分别=0.006和0.000）,MAP组与健康对照组间则差异无统计学意义（P=0.322）.以SAP组与MAP组血浆DNA含量绘制的ROC曲线下面积为0.881（95%可信区间为0.773～0.989）.以血浆DNA≥11.20 ng/ml为临界值,以血浆DNA含量诊断SAP的敏感度为19/20%,特异度为72.5%.血浆DNA含量与AP患者APACHE-Ⅱ评分、Ranson评分、血清Ca2＋、血清CRP正相关（P值分别=0.001、0.013、0.000、0.001）.结论 血浆DNA水平与AP病情严重程度相关,有可能成为AP病程监测及SAP早期诊断的重要指标.

Objective To quantitatively detect the circulating DNA in plasma of patients with acute pancreatitis and to evaluate its potential clinical values. Methods Blood samples collected from 40 patients with mild acute pancreatitis （MAP）, 20 with severe acute pancreatitis （SAP） and 50healthy controls were extracted for DNA using genomic DNA extraction kit. The DNA level was detected by real-time fluorescence quantitative PCR assay. The clinical association of DNA level with acute pancreatitis was analyzed. The receiver operating characteristic （ROC） curve was set up. Results The median concentration of plasma DNA in SAP group was 24.84 ng/ml, which was significantly higher than that in MAP group （7.60 ng/ml, P=0.006） and the healthy controls （5.23 ng/ml, P=0.000）. The median concentration of plasma DNA in patients with acute pancreatitis was higher than in healthy controls （P=0.006）, however, there was no significant difference between MAP group and healthy controls （P=0.322）.The area under the ROC curve performed by the plasma DNA from SAP and MAP groups was 0. 881（95% CI, 0.773-0.989）. With a cutoff value of 11.20 ng/ml, the overall sensitivity was 19/20% and specificity was 72.5%. Plasma DNA level was found to be associated with APACHE-Ⅱ score （P=0.001）, Ranson score （P=0. 013）, serum Ca2＋ level （P=0.000）, and c-reactive protein levels （P=0.001）. Conclusions Plasma DNA is correlated with the extent of pancreatitis. It can be used in monitoring the development of acute pancreatitis and may be a potential marker for early diagnosis of SAP.