RNA-Stabilized Whole Blood Samples but Not Peripheral Blood Mononuclear Cells Can Be Stored for Prolonged Time Periods Prior to Transcriptome Analysis
RNA-Stabilized Whole Blood Samples but Not Peripheral Blood Mononuclear Cells Can Be Stored for Prolonged Time Periods Prior to Transcriptome Analysis
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DOI：
10.1016/j.jmoldx.2011.03.006
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发表时间：
2011-07-01
影响因子：
4.1
通讯作者：
Staratschek-Jox, Andrea
中科院分区：
文献类型：
--
作者：
Debey-Pascher, Svenja;Hofmann, Andrea;Staratschek-Jox, Andrea
基于微阵列的外周血作为替代组织的转录组分析已成为临床实施中的重要方法。但是，在常规临床环境中应用基因表达分析需要仔细考虑样品处理和RNA分离方法对基因表达谱结果的影响。我们评估了不同样品保存策略的影响（例如，外周血单核细胞的冷冻保存或paxgene稳定的全血样品的冷冻保存）对基因表达谱。从冷冻保存的外周血单核细胞中获得的表达谱与非毛状对应物样品的表达谱有很大不同。此外，发现冷冻保存的外周血单核细胞样品中的表达谱会随着储存期的增加而发生重大变化，而长期冻结Paxgene RNA稳定的全血样品并未显着影响基因表达谱的稳定性。本报告描述了重要的技术方面，促进了使用外周血建立强大而可靠的基因表达研究指导，并为在常规处理诊断目的进行可靠实施的策略提供了有前途的策略。 （J Mol诊断2011，13：452-460。doi：10.1016/j.jmoldx.2011.03.006）
Microarray-based transcriptome analysis of peripheral blood as surrogate tissue has become an important approach in clinical implementations. However, application of gene expression profiling in routine clinical settings requires careful consideration of the influence of sample handling and RNA isolation methods on gene expression profile outcome. We evaluated the effect of different sample preservation strategies (eg, cryopreservation of peripheral blood mononuclear cells or freezing of PAXgene-stabilized whole blood samples) on gene expression profiles. Expression profiles obtained from cryopreserved peripheral blood mononuclear cells differed substantially from those of their nonfrozen counterpart samples. Furthermore, expression profiles in cryopreserved peripheral blood mononuclear cell samples were found to undergo significant alterations with increasing storage period, whereas long-term freezing of PAXgene RNA stabilized whole blood samples did not significantly affect stability of gene expression profiles. This report describes important technical aspects contributing toward the establishment of robust and reliable guidance for gene expression studies using peripheral blood and provides a promising strategy for reliable implementation in routine handling for diagnostic purposes. (J Mol Diagn 2011, 13:452-460. DOI: 10.1016/j.jmoldx.2011.03.006)