国产和进口荧光定量PCR试剂检测HBV-DNA的比对分析  

Comparative analysis on domestic and imported fluorescent quantitative PCR reagents for detecting HBV-DNA

作　　者：张玥[1] 田文君[1] 刘义庆[1] 张炳昌[1] 张庆[1] 渠滕 刘春梅[1] 

机构地区：[1]山东大学附属省立医院检验科,济南250021

出　　处：《检验医学与临床》2015年第2期147-148,150,共3页Laboratory Medicine and Clinic

基　　金：国家自然科学基金资助项目(81102220);山东省自然科学基金资助项目(ZR2011HM019);山东省临床重点专科建设项目(鲁卫医字[2013]26号)

摘　　要：目的分析国产和进口实时荧光定量聚合酶链反应（PCR）试剂检测乙型肝炎病毒（HBV）-DNA的相关性,探讨国产和进口试剂在临床应用中的差异。方法使用国产和进口试剂平行检测262例乙型肝炎（乙肝）患者血浆当中的HBV-DNA水平,国产试剂采用手工提取标本,罗氏LightCycler480Ⅱ（LC480Ⅱ）进行核酸扩增;进口试剂则采用罗氏COBAS AmpliPrep（CAP）和COBAS Taqman48（CTM）进行标本提取和扩增;对HBV-DNA病毒载量数据进行对数转换（log10）,并将两组数据进行相关分析。结果国产和进口试剂检测结果,经线性拟合,R2=0.814 3。HBV-DNA浓度在10-10^3 IU/mL的标本,R2=0.300 6;浓度在104-10^5 IU/mL的标本,R^2=0.411 8;浓度在10^6-10^8 IU/mL的标本,R^2=0.801 7。进口试剂阳性检出率为75.57%（198/262）,明显高于国产试剂阳性检出率[65.65%（172/262）]。结论国产试剂和进口试剂相比,相关性良好。HBV-DNA浓度在10^6-10^8IU/mL时,相关性最高;浓度在10^4-10^5 IU/mL时,相关性一般;浓度在10-10^3 IU/mL时相关性较差,国产试剂与进口试剂有明显差异,灵敏度有待进一步提高。Objective To analyze the correlation between the domestic and imported real‐time fluorescent quantitative PCR reagents for detecting HBV‐DNA and to explore their difference in clinical application .Methods The domestic and imported reagents were used to parallelly detect the plasma HBV‐DNA content in 262 cases of hep‐atitis B .The domestic reagent adopted the sample extracting by adopting the manual method and the amplification for nucleic acid was performed by the Roche LightCycler480 Ⅱ （LC480 Ⅱ ）;the imported reagent used the Roche CO‐BAS AmpliPrep （CAP） and COBAS Taqman48（CTM ） for conducting the sample extracting and amplification;the HBV DNA viral load data （log10） was performed the logarithmic transformation and the 2 sets of data were conduc‐ted the correlation analysis .Results The detection results of the domestic and imported reagents were performed the linear fitting ,R^2 =0 .814 3 .The sample with the concentration range of 10 -10^3 IU/mL ,R2 = 0 .300 6 ;the sample with the concentration range of 10^4 -10^5 IU/mL ,R2 = 0 .411 8 ;the sample with the concentration range of 10^6 -10^8 IU/mL ,R^2 =0 .801 7 .The positive detection rate of the imported reagent was 75 .57% （198/262） ,which was signifi‐cantly higher than 65 .65% （172/262） of the domestic reagent .Conclusion There is a good correlation between the domestic reagent and imported reagent .The correlation is the highest when the HBV‐DNA concentration in the range of 106 -108 IU/mL ;the correlation is general when the concentration in the range of 10^4 -10^5 IU/mL ;the correlation is lowest when the concentration in the range of 10-10^3 IU/mL ,there is a significant difference between the domes‐tic reagent and imported reagent and the sensitivity of the domestic reagent remains to be further improved .

关 键 词：乙型肝炎病毒DNA 实时荧光定量PCR 试剂比对 血浆 

分 类 号：R512.91[医药卫生—内科学]