以硅类作为吸附材料的两种不同核酸提取方法在HCV-RNA定量检测中的应用比较  

Comparison in application of two different nucleic acid extraction methods with silicon as adsorbent in HCV-RNA quantitative detection

作　　者：徐琼[1] 刘桂香[1] 田登[1] 高玉清[1] 卢囡囡 XU Qiong;LIU Gui-xiang;TIAN Deng;GAO Yu-qing;LU Nan-nan(Qinghai Center for Disease Control and Prevention,Xining810007,China)

机构地区：[1]青海省疾病预防控制中心,西宁810007

出　　处：《医学动物防制》2021年第3期240-242,共3页Journal of Medical Pest Control

基　　金：国家科技重大专项-艾滋病和病毒性肝炎等重大传染病防治(2017ZX10103006)。

摘　　要：目的比较及评价以硅类作为吸附材料的两种不同核酸的提取方法在丙型病毒性肝炎定量检测中的效果。方法选取124份抗-HCV阳性血清,分别采用核酸纯化柱提取法和二氧化硅吸附法提取HCV-RNA,用实时荧光定量PCR对其提取效果进行比较分析。结果 (1)124份抗-HCV阳性血清中以柱提法提取核酸后进行HCV-RNA定量检测的阳性率为70.97%(88/124)。二氧化硅吸附法提取核酸后进行HCV-RNA定量检测的阳性率为58.06%(72/124)。(2)两种方法提取核酸浓度分组比较结果,病毒载量拷贝数为1.0×10^(6)和1.0×10^(7)时,两种方法检出份数相同,而拷贝数为1.0×10^(3)、1.0×10^(4)和1.0×10^(5)时,检出份数均不一致。结论采用核酸提取柱提取血清标本优越于二氧化硅吸附法,其中两种方法检出份数差异集中在病毒载量低浓度区间,RNA病毒低复制时,柱提法检出份数高于二氧化硅吸附法。Objective To compare and evaluate the effects of two different nucleic acid extraction methods with silicon as the adsorbent in hepatitis C virus quantitative detection.Methods One hundred and twenty four anti-HCV positive sera were selected,and HCV-RNA was respectively extracted by nucleic acid purification column extraction method and silica adsorption method,and their extraction effects were compared and analyzed by Real-time fluorescence quantitative PCR.Results(1)The positive rate of HCV-RNA quantitative detection in 124 anti-HCV positive sera after nucleic acid extraction by column extraction method was 70.97%(88/124).The positive rate of HCV-RNA quantitative detetion by silica adsorption method was 58.06%(72/124).(2)The nucleic acid concentration grouping comparison results of two methods showed that the number of cases detected by the two methods remained to be equivalent when the viral load copy number was 1.0×10^(6) and 1.0×10^(7) respectively,but,the number of detected cases wasn’t equivalentwhen the number of copies was 1.0×10^(3),1.0×10^(4) and 1.0×10^(5).Conclusion The nucleic acid extraction column used to extract serum samples better than silica adsorption method,the difference in the number of cases detected by two methods is concentrated in the low concentration range of viral load.The number of cases detected by column extraction was higher than that by silica adsorption when RNA virus replication was low.

关 键 词：丙肝病毒 核酸提取方法 实时荧光定量PCR 血清处理 聚合酶联反应 

分 类 号：R512.63[医药卫生—内科学]