Human blood RNA stabilization in samples collected and transported for a large biobank.
Human blood RNA stabilization in samples collected and transported for a large biobank.
复制标题
DOI：
10.1186/1756-0500-5-510
复制
发表时间：
2012-09-18
影响因子：
1.8
通讯作者：
Lipkin WI
中科院分区：
文献类型：
--
作者：
Duale N;Brunborg G;Rønningen KS;Briese T;Aarem J;Aas KK;Magnus P;Stoltenberg C;Susser E;Lipkin WI
挪威母亲和儿童队列研究（MoBa）是1999年启动的一项全国性的以人口为基础的妊娠队列研究，包括1999年至2008年期间招募的108,000多名孕妇。在本研究中，我们评估了将RNA分析整合到现有MoBa协议中的可行性。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-并评估了采集管中不理想血容量和通过标准邮件运输血液样本的影响。表征样品的终点是RNA质量和产量，以及所选基因的RNA转录稳定性。PAXgene和Tempus试管稳定的血液样品中均可提取高质量的RNA。从Tempus试管中收集的血液样本中获得的RNA产量始终高于PAXgene试管。从脐带血中获得的RNA产量（3 - 4倍）比两种类型的成人血液高。通过标准邮件运输样品对RNA质量和RNA转录稳定性的影响中等；运输样品的整体RNA质量较高。一些无法解释的基因表达变化被注意到，这似乎与从试管中收集的次优血容量有关。运输过程中的温度变化也很重要。我们的结果强烈表明，特殊的收集管对于RNA稳定是必要的，它们应该用于建立新的生物库。我们还表明，MoBa生物库中收集的50,000个样本提供了高质量和足够数量的RNA，可以进行基因表达分析，研究疾病与基因表达模式改变的关联。
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. 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. 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.