摘要
目的评价基于NP968核酸提取仪（磁珠法）的实时荧光定量PCR系统检测乙型肝炎病毒DNA（HBVDNA）的性能。方法采用NP968自动核酸提取仪（磁珠法）提取血浆HBVDNA,经实时荧光定量PCR系统检测,分析其精密度、线性、回收率、最低检出限、参考区间,并将检测结果与传统煮沸法进行相关性分析,从而评价该法对HBVDNA检测的性能。结果基于NP968自动核酸提取仪（磁珠法）的实时荧光定量PCR系统检测HBVDNA,其低、高水平批内精密度均〈3/5TEa（TEa=0.4/靶值×100%）,批间精密度均〈4/5TEa;在4.16E2-4.16E8 IU/ml范围内具良好线性关系（Y=1.010X＋0.271,R2=0.989）;回收率为104%;最低检出限1.00E2 IU/ml;参考区间为〈1.00E3 IU/ml;检测结果与煮沸法之间具良好相关（Y=0.995X＋0.526,R2=0.951）。结论基于NP968自动核酸提取仪（磁珠法）的实时荧光定量PCR系统实现了快速、自动化提取HBVDNA,对检测HBVDNA具良好检测性能,适合临床推广使用。
Objective To evaluate the performance of NP968 nucleic acid extraction（MACS）-based real-time fluorescence quantitative PCR detection system for hepatitis B virus DNA（HBV DNA）. Methods Using the NP968 automated nucleic acid extraction system（MACS） to extract HBV DNA from the serum, and the DNA was detected by real-time fluorescence quantitative PCR system. In order to evaluate the performance of the detection system for HBV DNA, the precision, linearity, recovery rate, the lowest detection limit and reference interval of the testing system have been analyzed, and the correlation analysis with traditional boiling method was carried out. Results In the NP968 nucleic acid extraction（MACS）-based real-time fluorescence quantitative PCR detection system for HBV DNA, the within-run precision of the low and high level are all 3/5TEa（target TEa=0.4/ value x100%） and the between-run precision are all 4/5TEa; In the range of 4.16E2-4.16E8 IU/ml,there has good linear relationship（Y =1.010 X ＋0.271, R2=0.989）; the recovery rate is 104%. The minimum detection limit is 1.00E2 IU/ml; reference interval is 1.00E3 IU/ml, and the detection results between boiling method and MACS-based real-time fluorescence quantitative PCR has a good correlation（X＋Y=0.995 0.995, R2=0.951）. Conclusion The NP968 nucleic acid extraction（MACS）-based real-time fluorescence quantitative PCR detection system can extract the HBV DNA quickly and automatically and has good detection performance,so the detection system is suitable for clinical application.
出处
《实验与检验医学》
CAS
2015年第5期563-567,共5页
Experimental and Laboratory Medicine
基金
福建省卫生系统中青年骨干人才培养项目(编号:2013-ZQN-JC-2)
