两种核酸提取方法对丙型肝炎病毒RNA检测效果的比较及应用评价

Comparison of effects between two kinds of nucleic acid extraction method for detecting HCV RNA and their application evaluation

目的比较两种核酸提取方法对实时荧光定量聚合酶链反应(FQ-PCR)检测丙型肝炎病毒(HCV)RNA病毒载量的影响。方法选择89例2012年10月至2013年6月在肝病中心住院的慢性丙型肝炎抗-HCV阳性患者,分别用磁珠法和TRIZOL法提取血清标本中HCV RNA,用FQ-PCR技术检测HCV RNA,并对结果进行对比分析,统计学处理采用χ2检验和t检验。结果 89例抗HCV阳性标本中以磁珠法提取核酸进行HCV RNA定量检测的阳性率为73.0%(65/89),以TRIZOL法提取核酸进行HCV RNA定量检测的阳性率为65.1%(58/89),两法提取方法检测阳性率比较有统计学意义(χ2=3.76,P<0.05)。两法所测浓度结果换算成对数值,经t检验两者差异无统计学意义(t=0.32,P>0.05)。结论两种方法均可获得较高的回收率、有较高的敏感度和特异度,磁珠法提取HCV RNA可用于HCV感染者的临床诊断和疗效观察。

Objective To compare the influences of two kinds of nucleic acid extraction method for detecting hepatitis C virus（HCV） RNA viral load by the real‐time fluorescenct quantitative（FQ） PCR .Methods 89 inpatients with chronic hepatitis C （CHC） and anti‐HCV positive in the Therapy Center for Liver Diseases of our hospital from October 2012 to June 2013 were selected and HCVRNA was extracted from the serum specimens by using the mag‐netic beads and Trizol methods .FQ‐PCR was adopted for detecting serum HCV RNA .The detected results were per‐formed the comparative analysis .The statistic processing adopted the χ^2 test and t test .Results The positive rate of serum HCV RNA extracted by the magnetic beads and Trizol methods were 73 .0% （65/89） and 65 .1% （58/89） re‐spectively ,and the difference between the two methods showed the statistical significance （χ^2 =3 .76 ,P〈0 .05） .By converting the concentration results detected by these two methods into the log values ,there was no statistically sig‐nificant difference between the two methods（t=0 .32 ,P〉0 .05） .Conclusion The two kinds of method can obtain the higher recovery rate with higher sensitivity and higher specificity .The magnetic beads method for extracting HCV RNA could be used in the clinical diagnosis and therapeutic observation in the HCV infected persons .