Department of Biomedical, Experimental and Clinical Sciences, University of Florence, Italy.
Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, Italy.
Clin Chim Acta. 2015 Feb 2;440:205-10. doi: 10.1016/j.cca.2014.12.004. Epub 2014 Dec 6.
DNA integrity is a critical part of the definition of genomic DNA (gDNA) quality and can influence downstream molecular applications. Pre-analytical variables as sample storage and DNA extraction methods can influence the quality and quantity of isolated DNA and affect molecular test performances. The aim of this paper is to investigate the role of blood sample storage and DNA extraction procedures on gDNA integrity and gDNA fragmentation impact on a molecular test.
157 DNA samples deriving from the Pan European 1st SPIDIA DNA External Quality Assessment (EQA), aimed to investigate the influence of blood storage on gDNA quality and quantity, have been analyzed by Pulsed Field Gel Electrophoresis and ImageJ imaging software. 157 DNA samples derived from the Pan European 1st SPIDIA DNA External Quality Assessment (EQA), which aimed to investigate the influence of blood storage on gDNA quality and quantity, have been analyzed by Pulsed Field Gel Electrophoresis and ImageJ imaging software.
RESULTS/CONCLUSIONS: Our results demonstrate that blood sample storage and DNA extraction procedures influence gDNA integrity and that the same blood sample which underwent a long range multiplex PCR based analytical test can provide different results if the adopted pre-analytical procedures are not standardized.
DNA 完整性是基因组 DNA(gDNA)质量定义的关键部分,会影响下游分子应用。样本储存和 DNA 提取方法等分析前变量会影响分离 DNA 的质量和数量,并影响分子检测的性能。本文旨在研究血液样本储存和 DNA 提取程序对 gDNA 完整性的影响,以及 gDNA 片段化对分子检测的影响。
通过脉冲场凝胶电泳和 ImageJ 成像软件分析了源自泛欧 1 期 SPIDIA DNA 外部质量评估(EQA)的 157 个 DNA 样本,旨在研究血液储存对 gDNA 质量和数量的影响。
结果/结论:我们的结果表明,血液样本储存和 DNA 提取程序会影响 gDNA 的完整性,如果不规范采用的分析前程序,即使是经过长范围多重 PCR 分析测试的同一血液样本,也可能会提供不同的结果。