Department of Wildlife Diseases, Leibniz Institute for Zoo and Wildlife Research (IZW), Berlin, Germany.
Programa de Pós-Graduação em Saúde e Produção Animal na Amazônia, Universidade Federal Rural da Amazônia (UFRA), Belém, Pará, Brazil.
Mol Ecol Resour. 2024 Aug;24(6):e13979. doi: 10.1111/1755-0998.13979. Epub 2024 May 23.
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.
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，具有挑战性。在许多情况下，血液样本用于基于核酸的物种鉴定、基因组学或病原体研究。在大多数情况下，保持冷链是不可能的，样本在受控储存条件可用之前，会在环境温度下长时间保存。干血斑（DBS）储存，即储存在纤维素基纸上的血液，已经广泛应用于数十年的野外样本采集和保存。然而，在这种基质上长期储存会如何影响核酸浓度和完整性尚不清楚。我们使用四种核酸提取方法，分析了在 Whatman 滤纸条 3 和 FTA 卡上储存长达 15 年的 DBS 与冷链储存样本的核酸质量。我们使用靶向杂交捕获法，从亚马逊森林中 12 只自由活动的灵长类动物的样本中检测病毒序列的能力，并确定是否可以提取线粒体基因组。结果表明，即使经过长时间储存，DBS 仍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。
