基于自动化提取HCV RNA荧光定量检测系统的性能验证  

Validation of the performance of a fluorescence quantitative detection system of HCV RNA based on automated extraction

作　　者：邓波[1] 何彪[1] 廖亚龙 黄革[2] DENG Bo;HE Biao;LIAO Ya-long;HUANG Ge(Laboratory of the Gaozhou People’s Hospital,Gaozhou 525200,China;Laboratory of Guangdong Provincial People’s Hospital(Guangdong Academy of Medical Sciences),Southern Medical University,Guangzhou 510000,China)

机构地区：[1]高州市人民医院检验科,广东高州525200 [2]南方医科大学附属广东省人民医院(广东省医学科学院)检验科,广东广州510000

出　　处：《广东医科大学学报》2024年第1期73-77,共5页Journal of Guangdong Medical University

摘　　要：目的 基于Stream SP96全自动核酸提取仪、ABI7500实时荧光定量PCR仪对丙型肝炎病毒(HCV)RNA定量检测系统的性能指标进行验证。方法 依据《临床实验室对商品定量试剂盒分析性能的验证》WS/T 420-2013性能验证方案,采用Stream SP96全自动核酸提取平台及ABI7500荧光定量PCR仪检测HCV RNA,对其准确度、精密度、线性区间、检测限、定量限和抗干扰能力等进行方法学性能验证。结果 低、高浓度标准物质的均值与靶值的误差分别为0.20和0.25,均小于靶值对数值±0.4 log;低浓度样本的批内、批间不精密度变异系数值分别为0.79%、1.01%,高浓度样本的批内、批间不精密度变异系数值分别为0.52%、1.22%,均<5%;线性相关系数r>0.980,线性区间可达20~1.0×10^(8),呈良好线性(R^(2)=0.997 3);检出限为20 IU/mL,最低定量限为50 IU/mL;含胆红素(300 mg/L)、血红蛋白(300 g/L)、甘油三酯(3 000 mg/L)的干扰物质对样本检测结果无影响。结论 HCV RNA实时荧光PCR定量法检测系统准确度、精密度、线性范围、检出限、定量限、抗干扰能力均符合厂家声明,能够满足临床对HCV RNA定量检测的需求。Objective To verify the performance indicators of HCV RNA quantitative detection system based on Stream SP96 Automated Nucleic Acid Extractor and ABI 7500 Real-Time PCR System.Methods According to the WS/T 420-2013 Performance Verification Scheme in the Verification of Analytical Performance of Commercial Quantitative Kits by Clinical Laboratory,the Stream SP96 Stream SP96 Automated Nucleic Acid Extractor and ABI 7500 Real-Time PCR System were used to test HCV RNA,and methodological performance verification was performed on the accuracy,precision,linear interval,limit of detection,quantitative limit and anti-interference ability.Results The errors between the mean and target values of the low and high-concentration reference materials were 0.20 and 0.25,respectively,which were less than the logarithmic value of the target value±0.4 log.The intra-batch and inter-batch precision coefficient of variation(CV)values for low-concentration specimens were 0.79%and 1.01%,respectively,and those for high-concentration specimens were 0.52%and 1.22%,respectively,which were all<5%.The linear correlation coefficient r>0.980,and the linear interval was up to 20-1.0×10^(8),showing good linearity(R^(2)=0.9973).The limit of detection was 20 IU/mL and the lowest limit of quantitation was 50 IU/mL;The interfering substances containing bilirubin(300 mg/L),hemoglobin(300 g/L),and triglyceride(3000 mg/L)had no effect on the specimen detection results.Conclusion The HCV RNA real-time PCR quantitative testing systems have the accuracy,precision,linear range,limit of detection,limit of quantitation and anti-interference ability in line with the manufacturer’s statement,and they can fully meet the clinical requirements for HCV RNA quantitative testing.

关 键 词：HCV RNA 荧光法 性能验证 

分 类 号：R440[医药卫生—诊断学]