Department of Laboratory Medicine, Yonsei University College of Medicine, Seoul, Korea.
Yonsei Med J. 2010 Jan;51(1):104-10. doi: 10.3349/ymj.2010.51.1.104. Epub 2009 Dec 29.
The extraction of nucleic acid is initially a limiting step for successful molecular-based diagnostic workup. This study aims to compare the effectiveness of three automated DNA extraction systems for clinical laboratory use.
Venous blood samples from 22 healthy volunteers were analyzed using QIAamp Blood Mini Kit (Qiagen), MagNA Pure LC Nucleic Acid Isolation Kit I (Roche), and Magtration-Magnazorb DNA common kit-200N (PSS). The concentration of extracted DNAs was measured by NanoDrop ND-1000 (PeqLab). Also, extracted DNAs were confirmed by applying in direct agarose gel electrophoresis and were amplified by polymerase chain reaction (PCR) for human beta-globin gene.
The corrected concentrations of extracted DNAs were 25.42 +/- 8.82 ng/microL (13.49-52.85 ng/microL) by QIAamp Blood Mini Kit (Qiagen), and 22.65 +/- 14.49 ng/microL (19.18-93.39 ng/microL) by MagNA Pure LC Nucleic Acid Isolation Kit I, and 22.35 +/- 6.47 ng/microL (12.57-35.08 ng/microL) by Magtration-Magnazorb DNA common kit-200N (PSS). No statistically significant difference was noticed among the three commercial kits (p > 0.05). Only the mean value of DNA purity through PSS was slightly lower than others. All the extracted DNAs were successfully identified in direct agarose gel electrophoresis. And all the product of beta-globin gene PCR showed a reproducible pattern of bands.
The effectiveness of the three automated extraction systems is of an equivalent level and good enough to produce reasonable results. Each laboratory could select the automated system according to its clinical and laboratory conditions.
核酸提取最初是成功进行基于分子的诊断工作的限制步骤。本研究旨在比较三种用于临床实验室的自动化 DNA 提取系统的效果。
对 22 名健康志愿者的静脉血样本使用 QIAamp Blood Mini Kit(Qiagen)、MagNA Pure LC Nucleic Acid Isolation Kit I(Roche)和 Magtration-Magnazorb DNA common kit-200N(PSS)进行分析。通过 NanoDrop ND-1000(PeqLab)测量提取的 DNA 浓度。此外,通过直接琼脂糖凝胶电泳应用和聚合酶链反应(PCR)扩增人类β-球蛋白基因来确认提取的 DNA。
使用 QIAamp Blood Mini Kit(Qiagen)提取的 DNA 浓度校正值为 25.42±8.82ng/μL(13.49-52.85ng/μL),使用 MagNA Pure LC Nucleic Acid Isolation Kit I 提取的 DNA 浓度校正值为 22.65±14.49ng/μL(19.18-93.39ng/μL),使用 Magtration-Magnazorb DNA common kit-200N(PSS)提取的 DNA 浓度校正值为 22.35±6.47ng/μL(12.57-35.08ng/μL)。三种商业试剂盒之间未观察到统计学上的显著差异(p>0.05)。仅通过 PSS 的 DNA 纯度平均值略低于其他试剂盒。直接琼脂糖凝胶电泳可成功鉴定所有提取的 DNA。所有β-球蛋白基因 PCR 的产物均显示出可重复的条带模式。
三种自动化提取系统的效果相当,足以产生合理的结果。每个实验室可以根据其临床和实验室条件选择自动化系统。