两种核酸提取方法对HCV-RNA检验结果的影响  

Effect of Two Nucleic Acid Extraction Methods HCV-RNA Test Results

作　　者：何晓清[1] 严伟泉 王建[1] 

机构地区：[1]广东省惠州市第三人民医院检验科,惠州516002

出　　处：《中国医药导刊》2015年第2期184-186,共3页Chinese Journal of Medicinal Guide

摘　　要：目的:讨论固相吸附法和氯仿-异丙醇核酸提取方法对丙型肝炎病毒-RNA检验结果的影响。方法:选取106例丙型肝炎患者,随机分为实验组和对照组两组,采集所有患者血液,分离血清。实验组的53例患者,采用固相吸附法提取血清中的核酸。对照组的53例患者,采用氯仿-异丙醇法提取血清中的核酸。提取后的核酸,应用荧光定量PCR技术检测丙型肝炎病毒-RNA的含量,比较两种提取方法对丙型肝炎病毒-RNA检验结果的灵敏度、抗干扰能力和阳性率。结果:经研究比较发现:1对照组血清采用氯仿-异丙醇法提取核酸,稀释1×104倍后,荧光定量PCR可以检测,继续稀释则无法检测。实验组按照固相吸附法提取核酸,稀释1×105倍后,荧光定量PCR可以检测,继续稀释则低于检测限。2对照组核酸荧光定量PCR检测后,阳性35例,阳性率66.04%。实验组检测后阳性44例,阳性率83.02%。两组结果比较P<0.05,有显著性差异。3当血红蛋白浓度为5g/L时,两组抗干扰能力差别不大,P>0.05。当血红蛋白浓度为50g/L时,两组结果比较P<0.05。结论:与氯仿-异丙醇核酸提取方法相比,固相吸附法提取的丙型肝炎病毒-RNA灵敏度、阳性率更高,抗溶血干扰能力更好,具有一定优势。Objective： To discuss the solid phase assay and chloroform-isopropanol nucleic acid extraction methods affect HCV-RNA test results.Methods：106 cases of hepatitis C patients were randomly divided into two groups,experimental and control groups,collecting all the patient＇s blood serum was separated. 53 patients in the experimental group,the use of solid phase adsorption method to extract nucleic acids in serum.53 patients in the control group,using chloroform-isopropanol extraction of nucleic acids in serum.After nucleic acid extraction,the application of quantitative PCR technique to detect HCV-RNA levels,and compare the two extraction methods for HCV-RNA test results of sensitivity,anti-interference ability and positive rate.Results：The study compared that：1Control group,serum using chloroform-isopropanol extraction of nucleic acid,1×10^4-fold dilution,can be detected by fluorescence quantitative PCR,continue dilution can not be detected.Experimental groups according to the solid phase adsorption method to extract nucleic acids,1×10^5-fold dilution,can be detected by fluorescence quantitative PCR,continue dilution is below the detection limit.2Control group after nucleic acid fluorescence quantitative PCR positive in 35 cases,the positive rate of 66.04%.After the experimental group tested positive in 44 cases,the positive rate of 83.02%.Two sets of results compare, P〈0.05,significant difference.3When the hemoglobin concentration 5g/L,the two anti-jamming capability is not very different,P〈0.05.When the hemoglobin concentration was 50g/L,the result of the comparison groups, P〈0.05.Conclusion：Chloroform-isopropanol compared nucleic acid extraction method,solidphase extraction assay HCV-RNA sensitivity,positive rate higher,better anti-interference ability hemolysis,has certain advantages.

关 键 词：固相吸附法 氯仿-异丙醇法 丙型肝炎病毒-RNA 

分 类 号：R512.63[医药卫生—内科学]