DA3200-ABI7500全自动核酸提取平台高敏HCV RNA定量检测性能验证

Performance Verification of DA3200-ABI7500 Automatic Nucleic Acid Extraction Platform for High Sensitive HCV RNA Quantitative Detection

目的验证DA3200-ABI7500全自动核酸提取高敏检测HCV RNA方法性能。方法参考美国临床实验室标准化协会批准指南(CSLI-EP),采用DA3200全自动核酸提取平台及荧光定量PCR检测HCV RNA,评价检测方法的核酸提取纯度、精密度、正确度、线性范围、分析灵敏度、特异度和抗污染能力等性能指标。结果核酸提取后A260nm/A280nm比值为2.02。高、低两个浓度样本的批间精密度和批内精密度CV值均≤5%。正确度与抗干扰能力方面,标本实测值与靶值之间的偏离度≤±0.4lg。在13~1.3 E+06 IU/ml范围内,检测结果与靶值有良好的线性关系(Y=0.984X-0.186,r^2=0.998)。最低检出限为20 IU/ml的检出率为100%,且系统具有较强的特异度和抗污染能力。结论基于DA3200-ABI7500全自动核酸提取高敏检测HCV RNA各项分析性能指标均较好,高敏检测对治疗过程的监测及治疗终点判定具有重要意义。

Objective To evaluate the DA3200-ABI7500 automatic nucleic acid testing platform for highly sensitive HCV RNA quantitative detection.Methods Based on American standard CLSI-EP,the DA3200 automatic nucleic acid extraction platform and PCR analysis system was applied to high sensitivity detect HCV RNA,the precision,accuracy,linear range,detection ability and anti-pollution ability of this method was analyzed.Results In terms of nucleic acid extraction,the ratio of A260nm/A280nm was 2.02,and the intra-assay and inter-assay precision CV values of high and low concentration samples were both≤5%.In accuracy and anti-interference ability,the deviation between the measured value of the specimen and the target value was≤±0.4lg,and there was a good linear relationship between the test results and the target value in the range of 13~1.3 E+06 IU/ml(Y=0.984X-0.186,r^2=0.998).The detection rate of 20 IU/ml was 100%.Conclusion Based on the DA3200-ABI7500 automatic nucleic acid extraction platform,the high sensitivity HCV-RNA quantitative detection performance indicators were all good,and its high-sensitivity detection is of great significance for the monitoring of the treatment process and the determination of treatment endpoints.