摘要
目的对6个品牌全自动核酸提取仪进行性能验证,以期建立全自动核酸提取仪临床选择和评价的性能验证方案。方法采用阴性血清对乙型肝炎病毒核酸(HBV-DNA)血清标准物进行系列稀释,配制梯度浓度样品A-F。各仪器检测结果与罗氏系统比较,评价防污染能力、准确性、重复性、检测下限及线性相关性。用第三方扩增试剂盒对各仪器提取的核酸进行扩增检测,评价各系统开放性。结果通过防污染实验的5个品牌仪器进一步验证。配套系统中,品牌一、四与罗氏结果偏倚最小,准确性佳;品牌一、二对样品A(1×10^4IU/mL)与C(1×10^2IU/mL)的检测结果极差小,重复性佳;品牌一、五对样品F(1×10^1IU/mL)的检出率为100%;品牌三线性相关系数为0.998,相关性最佳。开放性实验的准确性、重复性和检测下限低于配套系统,其中品牌一在各检测参数中均显示最佳性能。结论配套系统中各品牌优势各异,而品牌一在开放性评价中优势明显,基本建立全自动核酸提取仪临床选择和评价的性能验证方案。
Objective To compare the performances of 6 brands of automated nucleic acid extraction platforms in order to establish a performance verification scheme for the selection and evaluation of automated nucleic acid extraction systems. Methods Serial dilutions of standard serum containing HBVDNA were mixed with HBV negative serum,and the concentrations of sample A-F were prepared. Then,samples were detected by each platform and their results were compared with Roches system to access the antiinterference feature,accuracy,reproducibility,limit of detection so as linear correlation. A third-party amplification kit was used to detect the extracted nucleic acids of each platform so as to evaluate the openness of detection systems. Results Subsequent tests continued when no cross-contamination was observed in 5 brands of platforms. In original systems,brand 1 and 4 had smaller biases and the accuracies were better. In reproducibility test,the ranges of brand 1 and 2 were the smallest. The detection rates of sample F(1×10~1 IU/mL) were 100% both in brand 1 and 5. Band 3 showed the best linear correlation, and the correlation coefficient was 0.998. In openness test,brand 1 showed the best performance. However,the accuracy,reproducibility and lowest limit of detection by using the third-party amplification reagents were all worse than those of the original systems. Conclusion Various brands of instruments in the original system showed different advantages,while brand 1 performed the best in the open system. The performance verification scheme for the clinical selection and evaluation of the automated nucleic acid extraction instrument can be established.
作者
戎国栋
赵鸿
吴蕾
黄珮珺
王芳
张燕
徐婷
RONG Guodong;ZHAO Hong;WU Lei;HUANG Peijun;WANG Fang;ZHANG Yan;XU Ting(Department of Laboratory Medicine,First Affiliated Hospital of Nanjing Medical University,Nanjing,Jiangsu,China,210029)
出处
《分子诊断与治疗杂志》
2019年第3期233-237,共5页
Journal of Molecular Diagnostics and Therapy
基金
江苏省实验诊断学重点实验室(ZDXKB2016005)
南京医科大学“十三五”教育研究课题(QN2017139)
关键词
荧光定量PCR
磁珠
HBV-DNA
性能验证
Fluorescence quantitative PCR
Magnetic beads
HBV-DNA
Performance verification
