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您的血液正在运输途中:关于运输时间和温度对稳定化全血中RNA降解的影响因素

Your Blood is Out for Delivery: Considerations of Shipping Time and Temperature on Degradation of RNA from Stabilized Whole Blood.

作者信息

Stefanovic Filip, Brown Lauren G, MacDonald James, Bammler Theo, Rinchai Darawan, Nguyen Serena, Zeng Yuting, Shinkawa Victoria, Adams Karen, Chausabel Damien, Berthier Erwin, Haack Amanda J, Theberge Ashleigh B

机构信息

Department of Chemistry, University of Washington, Seattle, Washington 98195, United States.

Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, Washington 98195, United States.

出版信息

bioRxiv. 2024 Aug 26:2024.08.24.609519. doi: 10.1101/2024.08.24.609519.

Abstract

Remote research studies are an invaluable tool for reaching populations in geographical regions with limited access to large medical centers or universities. To expand the remote study toolkit, we have previously developed homeRNA, which allows for at-home self-collection and stabilization of blood and demonstrated the feasibility of using homeRNA in high temperature climates. Here, we expand upon this work through a systematic study exploring the effects of high temperature on RNA integrity through in-lab and field experiments. Compared to the frozen controls (overall mean RIN of 8.2, = 8), samples kept at 37°C for 2, 4, and 8 days had mean RINs of 7.6, 5.9, and 5.2 ( = 3), respectively, indicating that typical shipping conditions (~2 days) yield samples suitable for downstream RNA sequencing. Shorter time intervals (6 hours) resulted in minimal RNA degradation (median RIN of 6.4, = 3) even at higher temperatures (50°C) compared to the frozen control (mean RIN of 7.8, = 3). Additionally, we shipped homeRNA-stabilized blood from a single donor to 14 different states and back during the summer with continuous temperature probes (7.1 median RIN, = 42). Samples from all locations were analyzed with 3' mRNA-seq to assess differences in gene counts, with the transcriptomic data suggesting that there was no preferential degradation of transcripts as a result of different shipping times, temperatures, and regions. Overall, our data support that homeRNA can be used in elevated temperature conditions, enabling decentralized sample collection for telemedicine, global health, and clinical research.

摘要

远程研究对于接触那些难以进入大型医疗中心或大学的地理区域的人群来说是一种非常宝贵的工具。为了扩展远程研究工具包,我们之前开发了homeRNA,它允许在家中自行采集和稳定血液样本,并证明了在高温气候下使用homeRNA的可行性。在这里,我们通过一项系统研究来扩展这项工作,该研究通过实验室和实地实验探索高温对RNA完整性的影响。与冷冻对照组(总体平均RIN为8.2,n = 8)相比,在37°C下保存2天、4天和8天的样本的平均RIN分别为7.6、5.9和5.2(n = 3),这表明典型的运输条件(约2天)产生的样本适合进行下游RNA测序。即使在较高温度(50°C)下,较短的时间间隔(6小时)导致的RNA降解也最小(中位数RIN为6.4,n = 3),而冷冻对照组的平均RIN为7.8(n = 3)。此外,我们在夏季使用连续温度探头将来自单个捐赠者的homeRNA稳定化血液运往14个不同的州并运回(中位数RIN为7.1,n = 42)。对所有地点的样本进行3' mRNA测序分析以评估基因计数的差异,转录组数据表明,由于不同的运输时间、温度和地区,转录本没有出现优先降解的情况。总体而言,我们的数据支持homeRNA可在高温条件下使用,从而能够为远程医疗、全球健康和临床研究进行分散式样本采集。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d610/11370555/50b64890ec37/nihpp-2024.08.24.609519v2-f0001.jpg

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