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在为大型生物样本库采集和运输的样本中实现人血RNA的稳定化。

Human blood RNA stabilization in samples collected and transported for a large biobank.

作者信息

Duale Nur, Brunborg Gunnar, Rønningen Kjersti S, Briese Thomas, Aarem Jeanette, Aas Kaja K, Magnus Per, Stoltenberg Camilla, Susser Ezra, Lipkin W Ian

机构信息

The Norwegian Institute of Public Health, Oslo, Norway.

出版信息

BMC Res Notes. 2012 Sep 18;5:510. doi: 10.1186/1756-0500-5-510.

Abstract

BACKGROUND

The Norwegian Mother and Child Cohort Study (MoBa) is a nation-wide population-based pregnancy cohort initiated in 1999, comprising more than 108.000 pregnancies recruited between 1999 and 2008. In this study we evaluated the feasibility of integrating RNA analyses into existing MoBa protocols. We compared two different blood RNA collection tube systems - the PAXgene™ Blood RNA system and the Tempus™ Blood RNA system - and assessed the effects of suboptimal blood volumes in collection tubes and of transportation of blood samples by standard mail. Endpoints to characterize the samples were RNA quality and yield, and the RNA transcript stability of selected genes.

FINDINGS

High-quality RNA could be extracted from blood samples stabilized with both PAXgene and Tempus tubes. The RNA yields obtained from the blood samples collected in Tempus tubes were consistently higher than from PAXgene tubes. Higher RNA yields were obtained from cord blood (3 - 4 times) compared to adult blood with both types of tubes. Transportation of samples by standard mail had moderate effects on RNA quality and RNA transcript stability; the overall RNA quality of the transported samples was high. Some unexplained changes in gene expression were noted, which seemed to correlate with suboptimal blood volumes collected in the tubes. Temperature variations during transportation may also be of some importance.

CONCLUSIONS

Our results strongly suggest that special collection tubes are necessary for RNA stabilization and they should be used for establishing new biobanks. We also show that the 50,000 samples collected in the MoBa biobank provide RNA of high quality and in sufficient amounts to allow gene expression analyses for studying the association of disease with altered patterns of gene expression.

摘要

背景

挪威母婴队列研究(MoBa)是一项始于1999年的全国性基于人群的妊娠队列研究,包含1999年至2008年间招募的超过108,000例妊娠。在本研究中,我们评估了将RNA分析整合到现有MoBa方案中的可行性。我们比较了两种不同的血液RNA采集管系统——PAXgene™血液RNA系统和Tempus™血液RNA系统——并评估了采集管中血液量不足以及通过标准邮件运输血样的影响。表征样本的终点指标是RNA质量和产量,以及选定基因的RNA转录本稳定性。

研究结果

使用PAXgene管和Tempus管稳定的血样均可提取出高质量的RNA。从Tempus管采集的血样中获得的RNA产量始终高于PAXgene管。两种类型的管中,脐血的RNA产量均高于成人血液(3至4倍)。通过标准邮件运输样本对RNA质量和RNA转录本稳定性有中度影响;运输样本的总体RNA质量较高。注意到一些基因表达的不明原因变化,这似乎与采集管中血液量不足有关。运输过程中的温度变化可能也有一定影响。

结论

我们的结果强烈表明,RNA稳定需要特殊的采集管,并且应将其用于建立新的生物样本库。我们还表明,MoBa生物样本库中收集的50,000个样本提供了高质量且足够量的RNA,足以进行基因表达分析,以研究疾病与基因表达改变模式之间的关联。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/06e9/3503553/c285ce64d0a5/1756-0500-5-510-3.jpg

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