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 20～1.0×108,呈良好线性(R2=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.
Abstract
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 20-1.0×108,showing good linearity(R2=0.997 3).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(3 000 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.
维普
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