核酸纯化柱提取核酸定量检测丙型肝炎病毒RNA的临床应用

Clinical application of nucleic acid purification mini spin column quantitating HCV-RNA

目的观察核酸纯化柱提取法(简称柱提法)对血浆标本中丙型肝炎(简称丙肝)病毒核糖核酸(HCV-RNA)进行定量检测的临床应用效果。方法分别用柱提法和酚-氯仿提取法提取血浆标本中的HCV-RNA,用实时荧光定量聚合酶链反应(FQ-PCR)技术对117例抗-HCV阳性的丙肝患者同时进行HCV-RNA定量检测,并对结果进行比较分析。结果117例抗-HCV阳性丙肝患者以酚-氯仿提取法提取核酸进行HCV-RNA定量检测的阳性率为66.7%(78/117),以柱提法提取核酸进行HCV-RNA定量检测的阳性率为79.5%(93/117)。两法比较,差异有统计学意义(χ2=4.26,P<0.05)。将两法所测浓度结果换算成对数值,经u检验两者差异无统计学意义(u=1.61,P>0.05)。结论采用核酸柱提法提取血浆标本中HCV-RNA进行定量检测简单、快速,分离效率高,容易掌握,适于临床常规应用。

Objective To investigate the clinical application of nucleic acid purification mini spin column quan- titating HCV-RNA of plasma samples. Methods Nucleic acid purification mini spin column extraction assay and phenol-chloroform extractio were applied to extracting HCV-RNA of plasma samples, and then real-time fluorescent quantitation polymerase chain reaction （FQ-PCR） was adopted to detect plasma HCV-RNA in 117 patients with hep- atitis C （with anti-HCV positive）. The results were compared and analyzed. Results The positive rate of plasma HCV-RNA collected by phenol-chloroform extraction and by column extraction assay were 66. 7% （78/117） and 79.5% （93/117） respectively. There was statistical difference in the positive rate of plasma HCV-RNA beteen the two methods （X^2 =4.26,P〈0.05）. By comparison, there was not significant difference statistically in the copies of plasma HCV-RNA between the two groups （u= 1.61,P〉0.05）. Conclusion Plasma HCV-RNA extration by nucleic acid purification mini spin column extraction assay for quantitively determination is simple, rapid, wield and efficient, suitable for clinical routine application.