Taqman探针-荧光定量PCR法检测HBVDNA的性能验证及评价  

Performance Verification and Evaluation of Taqman Probe-Fluorescence Quantitative Polymerase Chain Reaction for Detection of HBV DNA

作　　者：孙慧[1] 叶光明[1] 黎安玲[1] 刘松梅[1] 王诗婵 SUN Hui;YE Guangming;LI Anling;LIU Songmei;WANG Shichan(Clinical Gene Amplification Laboratory,ZhongNan Hospital,Wuhan University,430071)

机构地区：[1]武汉大学中南医院检验科临床基因扩增检验室,430071

出　　处：《国际检验医学杂志》2019年第S02期39-43,共5页International Journal of Laboratory Medicine

摘　　要：目的探讨适合本实验室的Taqman探针-荧光定量PCR法检测HBV-DNA的性能验证方法,评估其分析性能及其临床应用价值。方法采用卫健委临检中心室间质评参考物质、第三方质控品、和武汉大学中南医院收集的临床标本,按方法学性能评价指标设计验证试验,对检测结果的精密度、正确度、灵敏度、线性范围和干扰因素等参数进行性能验证。结果本检测系统批内精密度和批间精密度CV均小于<5%;正确度符合卫健委临检中心室间质评要求;检测灵敏度可以达到50 IU/mL,CV值≤20%;在5.90E+1 IU/mL^6.34 E+7 IU/mL浓度范围内,线性回归方程为Y=0.9833 X+0.0732,R^2=0.999;当血红蛋白≤23.5 g/L、胆红素≤450μM、三酰甘油≤22.5 mM时,检测结果未受干扰。结论经验证,该荧光定量PCR检测系统检测HBV-DNA的性能指标满足临床预期要求,常规应用结果满意;荧光定量PCR的性能验证是保证其检验质量的必要手段,形成适合的验证方案流程将会对临床基因扩增检验的标准化和规范化起到重要作用。Objective To explore and evaluate the analytical performance and clinical application value of the validation method of Taqman probe-fluorescence quantitative PCR for detection of HBV-DNA in laboratory.Methods Analytical performance of the HBV DNA quantitative detecting reagents included precision,accuracy,functional sensitivity,linear range and interference factors were verified by collecting high concentration samples,third party quality products and reference materials from National Center for Clinical Laboratories(NCCL).Results The within-run coefficient of variation(CV)of both low and high concentration samples were below 5%,meanwhile the intra-assay CV was below 5%too.Detection sensitivity was able to attain 50 IU/mL.while the CV was below 20%.It exhibited a benign linear relation from 5.90E+1IU/mL to 6.34E+7IU/mL,Y=0.9833 X+0.0732,R^2=0.999.When hemoglobin≤23.5 g/L,bilirubin≤450μM and triglyceride≤22.5 mM,the results were not disturbed.Conclusion It has been proved that the performance of the fluorescent quantitative PCR system for detecting HBV-DNA meets the clinical expectations.The performance verification of fluorescence quantitative PCR is a necessary means to ensure the quality of the test.Forming a suitable validation program flow will play an important role in standardization and standardization of clinical gene amplification test.

关 键 词：乙型肝炎病毒 TAQMAN探针 荧光定量PCR 性能验证 

分 类 号：R44[医药卫生—诊断学]