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

摘要

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 (χ2 =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 .%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 HCV-RNA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。