Abstract:
Collecting and preserving biological samples in the field, particularly in remote areas in tropical forests, prior to laboratory analysis is challenging. Blood samples in many cases are used for nucleic acid-based species determination, genomics or pathogen research. In most cases, maintaining a cold chain is impossible and samples remain at ambient temperature for extended periods of time before controlled storage conditions become available. Dried blood spot (DBS) storage, blood stored on cellulose-based paper, has been widely applied to facilitate sample collection and preservation in the field for decades. However, it is unclear how long-term storage on this substrate affects nucleic acid concentration and integrity. We analysed nucleic acid quality from DBS stored on Whatman filter paper no. 3 and FTA cards for up to 15 years in comparison to cold-chain stored samples using four nucleic acid extraction methods. We examined the ability to identify viral sequences from samples of 12 free-ranging primates in the Amazon forest, using targeted hybridization capture, and determined if mitochondrial genomes could be retrieved. The results suggest that even after extended periods of storage, DBS will be suitable for some genomic applications but may be of limited use for viral pathogen research, particularly RNA viruses.
摘要:
在野外收集和保存生物样本,特别是在热带森林的偏远地区,在实验室分析之前是具有挑战性的。在许多情况下,血液样本用于基于核酸的物种测定,基因组学或病原体研究。在大多数情况下,维持冷链是不可能的,并且样品在受控储存条件变得可用之前在环境温度下保持延长的时间段。干血斑(DBS)储存,血液储存在纤维素基纸上,几十年来一直被广泛应用于方便样品的采集和保存领域。然而,目前尚不清楚在该底物上的长期储存如何影响核酸浓度和完整性。我们分析了保存在Whatman滤纸上的DBS的核酸质量。3和FTA卡长达15年,与使用四种核酸提取方法的冷链储存样品相比。我们检查了从亚马逊森林中12种自由放养的灵长类动物的样本中鉴定病毒序列的能力,使用靶向杂交捕获,并确定是否可以检索线粒体基因组。结果表明,即使经过长时间的储存,DBS将适用于某些基因组应用,但可能对病毒病原体研究的用途有限。特别是RNA病毒。