丙型肝炎病毒-核糖核酸定量检测系统的性能验证及评价  

Performance Verification and Evaluation of HCV-RNA Quantitative Detection System

作　　者：张彦懿 杨佳佳[1] 骆贞红 刘华伟[1] 黄江渝[1] Zhang Yanyi;Yang Jiajia;Luo Zhenhong;Liu Huawei;Huang Jiangyu(Department of Clinical Medical Laboratory, The Third People's Hospital of Chengdu, Chengdu 610031, China)

机构地区：[1]成都市第三人民医院临床医学检验部,成都610031

出　　处：《成都医学院学报》2020年第6期706-710,共5页Journal of Chengdu Medical College

基　　金：四川省医学会科研项目(No:S17010)。

摘　　要：目的对基于Smart 32核酸提取仪和Roche Cobas z480实时荧光PCR仪的丙型肝炎病毒(HCV)-核糖核酸(RNA)定量检测系统进行性能验证。方法依据中国合格评定国家认可委员会(CNAS)-GL039《分子诊断检验程序性能验证指南》的方案,采用标准物质及临床样本,对检测系统的正确度、精密度、线性范围、抗干扰能力、检出限、最低定量限进行验证,同时计算各性能参数与制造商声明比较。结果精密度验证结果显示,高、低浓度水平样本的重复性精密度变异系数(CV)值分别为1.54%、3.61%,中间精密度CV值分别为2.16%、4.42%,均小于厂家声明的5%。正确度:低、中值两个浓度水平标准物质的均值与靶值的误差分别为0.06和0.08,均<±0.4。线性范围:经拟合分析,证实在50~1.2×107 IU/mL呈良好线性(R2=0.9955>0.97)。抗干扰能力验证:结果显示,含30 mg/dL胆红素、3.2 g/dL甘油三酯、30 g/dL血红蛋白、6 g/dL白蛋白的干扰物质对样本检测结果无影响,且对甘油三酯和血红蛋白的抗干扰能力略高于厂家声明。检出限20 IU/mL,最低定量限50 IU/mL也与制造商声明一致。结论HCV-RNA定量检测系统正确度、精密度、线性范围、抗干扰能力、检出限、最低定量限各项性能指标与制造商声明一致,满足国际标准化组织(ISO)15189要求,可用于临床检测。Objective To verify the performance of hepatitis C virus(HCV)-ribonucleic acid(RNA)quantitative detection system based on Smart 32 nucleic acid extractor and Roche Cobas z480 real-time fluorescent PCR instrument.Methods According to China National Accreditation Service for Conformity Assessment(CNAS)-GL039 Guidelines for Performance Verification of Molecular Diagnostic Test Procedures,standard substances and clinical samples were used to verify the accuracy,precision,linear range,anti-interference ability,detection limit and minimum quantitative limit of the testing system.And the performance parameters were calculated and compared with the manufacturer's statement.Results Precision verification results showed that the repeatability precision coefficient of variation(CV)values of high and low concentration samples were 1.54%and 3.61%,respectively,and the intermediate precision CV values were 2.16%and 4.42%,respectively,which were all less than the 5%stated by the manufacturer.As to accuracy,the mean and target errors of standard substances at low and median concentration levels were 0.06 and 0.08 respectively,both were less than±0.4.Fitting analysis confirmed good linearity when the linear range was 50-1.2×107 IU/mL(R2=0.9955>0.97).Anti-interference ability verification results showed that the interferences of 30 mg/dL total bilirubin,3.2 g/dL triglyceride,30 g/dL hemoglobin,6 g/dL albumin had no influence on the test results of the samples.And the anti-interference ability to triglyceride and hemoglobin was slightly higher than the manufacturer's statement.The detection limit 20 IU/mL and the minimum quantitative limit 50 IU/mL were consistent with manufacturer's statement.Conclusion The accuracy,precision,linear range,anti-interference ability,detection limit and minimum quantitative limit of HCV-RNA quantitative detection system are consistent with the manufacturer's statement.This detection system meets the requirements of International Organization for Standardization(ISO)15189,and can be used for clinical

关 键 词：性能验证 丙型肝炎病毒 定量限 核酸定量 

分 类 号：R446.9[医药卫生—诊断学]