Nucleic acid degradation after long‐term dried blood spot storage

Author:

Li Juan1ORCID,Ulloa Gabriela M.23,Mayor Pedro245,Santolalla Robles Meddly L.6,Greenwood Alex D.17ORCID

Affiliation:

1. Department of Wildlife Diseases Leibniz Institute for Zoo and Wildlife Research (IZW) Berlin Germany

2. Programa de Pós‐Graduação em Saúde e Produção Animal na Amazônia Universidade Federal Rural da Amazônia (UFRA) Belém Pará Brazil

3. Grupo de Enfermedades Infecciosas Re‐emergentes Universidad Científica del Sur (UCSUR) Lima Peru

4. ComFauna, Comunidad de Manejo de Fauna Silvestre en la Amazonía y en Latinoamérica Iquitos Peru

5. Departament de Sanitat i Anatomia Animals Universitat Autònoma de Barcelona Bellaterra Spain

6. Emerge, Emerging Diseases and Climate Change Research Unit, School of Public Health and Administration Universidad Peruana Cayetano Heredia Lima Peru

7. School of Veterinary Medicine Freie Universität Berlin Berlin Germany

Abstract

AbstractCollecting 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.

Publisher

Wiley

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