磁珠法结合实时荧光PCR试剂检测HBV-DNA临床性能验证  

Clinical performance validation of Real-time fluorescence PCR for detection of HBV-DNA by magnetic beads extraction

作　　者：石莹[1] 田绿波[1] 樊学军[1] 王俊贤[1] 龙娟[2] SHI Ying;TIAN Lübo;FAN Xuejun;WANG Junxian;LONG Juan(Sichuan International Healthcare Center,Chengdu,Sichuan 610041,China;不详)

机构地区：[1]四川国际旅行卫生保健中心,四川成都610041 [2]四川省人民医院临床医学检验中心

出　　处：《中国国境卫生检疫杂志》2023年第5期454-459,共6页Chinese Journal of Frontier Health and Quarantine

基　　金：海关总署科研项目(2020HK119,2019HK135)。

摘　　要：目的 分析磁珠法提取核酸结合实时荧光定量PCR试剂检测HBV-DNA的性能及其临床应用价值,有助于选择试剂。方法 按照CNAS-CL36能力验证要求,收集四川省人民医院乙肝病人血清样本,对三家磁珠法核酸提取实时荧光定量PCR试剂(A、B、C试剂)的精密度、正确度、检出限、可报告范围、抗干扰能力等等性能参数进行方法学性能验证和评价。结果 试剂A和B均能检出HBV-DNA,检出率100%,试剂C的检出率为83.3%,与厂家公布的定量检测下限20 IU/ml不一致。试剂A检测低浓度和高浓度标本的批内变异系数(CV)均低于5%;试剂B检测低浓度标本的批间CV≥5%,高浓度标本批间CV≤5%,试剂C检测低浓度和高浓度标本的批间CV均≥5%。3种试剂在分析测量范围内均线性关系良好。干扰试验显示,3种试剂均受到严重溶血、脂血标本的影响,但不受轻微溶血、脂血标本的影响。结论 通过对3种试剂的性能验证,达安基因公司的一步法(磁珠法)HBV-DNA定量检测试剂灵敏度高,有利于乙肝筛查和临床疗效的监测、随访。Objective To evaluate the analytical performance and clinical value of Real-time fluorescent quantitative PCR reagents for HBV-DNA by nucleic acid magnetic beads extraction reagents,to help to select reagents.Methods According to the requirements of CNAS-CL36 capability verification,serum samples of hepatitis B patients from Sichuan Provincial People's Hospital were collected,and methodological performance of the three(reagents A,B and C) nucleic acid extraction Real-time fluorescent quantitative PCR reagents by magnetic beads were verified and evaluated,including precision,accuracy,detection limit,reporting range and anti-interference ability.Results The LOD of reagents A and B met the performance parameters,the positive detection rate were all 100%,and the positive detection rate of reagent C was 83.3%,which was inconsistent with the lower limit of quantitative detection20 IU/ml announced by the manufacturer.The coefficient of variation(CV) of reagent A low-concentration and high-concentration specimens were lower than 5% in batches,reagent B low-concentration specimens was CV≥5%,high-concentration specimens was CV ≤5%,reagent C low-concentration and high-concentration specimens were CV≥5%.There was good correlation between the results of 3 reagents in the linear range.The interference test of the three reagents showed that severe hemolysis and lipid samples had an effect on the detection of HBV-DNA with low viral load,but were not affected by mild hemolysis and lipid samples.Conclusion Through the performance verification of the three reagents,it was found that one-step(magnetic bead method) quantitative detection reagent for HBV-DNA made by Daan Gene Co.Ltd had high sensitivity,and suitable for port and clinical routine detection.

关 键 词：乙肝病毒 定量检测 性能验证 荧光定量PCR 

分 类 号：R185[医药卫生—流行病学]