HBV-DNA实时荧光定量PCR检测试剂性能评价

The performance evaluation of PCR detection reagents by real-time fluorescence quantification of HBV-DNA

目的评估实时荧光定量PCR法检测乙型肝炎病毒(HBV-DNA)核酸检测试剂的分析性能及结果的临床应用价值。方法采用河北省故城县医院收集的高浓度阳性标本和国家卫生部临检中心和河北省临检中心的室间质控品,对乙型肝炎病毒(HBV-DNA)检测试剂的精密度,正确度,检测限(功能灵敏度),分析测量范围等性能参数进行方法学性能验证和评估。结果高浓度和低浓度的批内精密度和批间精密度标准偏差均≤5%,正确度的检测结果符合国家卫生健康委临床检验中心和河北省临床检验的室间质评要求。检测下限(功能灵敏度)可达到200 U/ml,且重复间的CV值≤20%;在2.0E+02-1.0E+07分析测量范围线性关系良好(线性关系系数R2=0.99)。结论定量项目检测正式用于检测临床标本前必须对检测系统的分析性能做充分的评估。通过实验室验证HBV-DNA实时荧光定量PCR检测试剂可以满足目前乙肝的筛查和临床疗效监测的需求,并且检测过程经济简便,适用于临床的常规检测。

Objective To investigate and evaluate the performance of real-time fluorescence quantitative PCR detection system in detecting hepatitis B virus(HBV-DNA).Methods The positive specimens collected by Gucheng county hospital and quality control samples from clinical detection center of Ministry of Public Health were enrolled in the study.The precision,accuracy,detection limit(functional sensitivity)of the test reagents of detecting HBV-DNA were examined to evaluate the test performance.Results The standard deviations of within-run precision and between-run precision in high-concentration specimens and low-concentration specimens were less than 5%,and the accuracy test results were consistent with the related request.The lower limit of was 200U/ml,with CV value≤20%.The linear range was 2.0E+02-1.0E+07,and the R2 of linear evaluation was 0.99.Conclusion The evaluation results of the PCR detection system meet the requirements of the related documents for the performance evaluation of the nucleic acid detection system.The performance evaluation scheme of HBV detection system in the laboratory is feasible and practical,it can be used to evaluate the HBV-DNA quantitative detection system.