两种核酸提取方法检测血清丙型肝炎病毒核糖核酸的比较

Comparing of two methods of nucleic acid extraction for detecting hepatitis C virus RNA in serum

目的比较两种不同核酸提取方法检测血清丙型肝炎病毒核糖核酸(HCV-RNA)对临床诊断的应用价值。方法对128份HCV抗体阳性的患者血清,同时用两种实时荧光定量聚合酶链反应(PCR)方法检测HCV-RNA并检测丙氨酸氨基转移酶(ALT)水平,这两种方法分别是用二氧化硅微粒法提取RNA和纯化柱法提取RNA。结果 128份HCV抗体阳性的血清中二氧化硅微粒法HCV-RNA阳性率41.41%,纯化柱法HCVRNA阳性率59.38%,纯化柱法优于二氧化硅微粒法,差异有统计学意义(χ2=8.27,P<0.05)。49份HCV抗体阳性的血清中ALT异常,且ALT浓度变化与纯化柱法HCV-RNA水平呈正相关性(r=0.95,P<0.05)。结论血清HCV-RNA检测对丙型肝炎诊断和病情监测均有重要的临床意义,采用纯化柱法提取RNA具有更优的临床价值。

Objective To compare the value of two different methods of nucleic acid extraction for detecting serum hepatitis C RNA （HCV RNA） on the clinical diagnosis. Methods Two methods of real-time fluorescence quantitative PCR were applied to detect HCV-RNA in 128 HCV antibody-positive patients,and the alanine transarni nase （ALT） level was tested. These two methods were silica particles RNA extraction and RNA purified by column. Results In the 128 samples of HCV antibody-positive sera, HCV-RNA positive rate was 41.41% by silica particles RNA extraction, and the rate was 59.38% by using column purification RNA. Purification column method was better than silica particles,and there was significant difference in positive rate （X2 =8.27 ,P〈0.05）. In 49 samples of HCV antibody-positive sera, the ALT level was abnormal, and there was positive correlation between ALT level and HCV- RNA content detected by purification column （r= 0. 95,P〈0. 05）. Conclusion Detecting HCV-RNA in serum could be useful for clinical diagnose and therapy effects monitoring, and purification column method is better.