Translational Medicine Research Group, Cranfield Health, Cranfield University, Cranfield, United Kingdom.
J Immunol Methods. 2012 Oct 31;384(1-2):118-27. doi: 10.1016/j.jim.2012.07.016. Epub 2012 Jul 31.
Automated nucleic acid extractions from dried blood spot (DBS) samples promises standardized sample treatment, low error rates, avoidance of contamination and requirement of less hands-on time. In the present study, non-automated and automated column based extraction processes using the QIAamp Investigator procedure were compared for the extraction of DNA from DBS samples. The concentration and the purity of DNA generated were determined by optical density readings. Furthermore qPCR downstream applications using the nucleic acids extracted with the two processes and albumin and T-cell receptor excision circles (TREC) copy numbers were measured and compared. The influence of the time of storage was also investigated by analyzing samples freshly dried and stored up to 11weeks at -20°C from the same individual. Finally, we provide arguments of preferentially choosing the automated procedure for extracting DNAs from DBS samples when downstream qPCR applications are required.
从干血斑 (DBS) 样本中自动提取核酸有望实现标准化的样本处理、低错误率、避免污染和减少人工操作时间。在本研究中，比较了使用 QIAamp Investigator 程序的非自动化和自动化柱基提取过程，用于从 DBS 样本中提取 DNA。通过吸光度读数确定生成的 DNA 的浓度和纯度。此外，使用两种方法提取的核酸进行 qPCR 下游应用，并测量和比较白蛋白和 T 细胞受体切除环 (TREC) 的拷贝数。通过分析同一个体新干燥并在-20°C 下储存长达 11 周的样本，研究了储存时间的影响。最后，当需要进行 qPCR 下游应用时，我们提供了从 DBS 样本中提取 DNA 时优先选择自动化程序的论据。
