Successful restoration of archived ovine formalin fixed paraffin-embedded tissue DNA and single nucleotide polymorphism analysis.


Journal

Veterinary research communications
ISSN: 1573-7446
Titre abrégé: Vet Res Commun
Pays: Switzerland
ID NLM: 8100520

Informations de publication

Date de publication:
Jan 2023
Historique:
received: 23 03 2022
accepted: 09 05 2022
pubmed: 27 5 2022
medline: 27 1 2023
entrez: 26 5 2022
Statut: ppublish

Résumé

Archived formalin fixed paraffin-embedded (FFPE) tissues are powerful tools in medicine, capable of harboring diagnostic and genetic answers to challenging clinical questions. Successful utilization of DNA derived from FFPE samples is dependent upon repairing DNA damage generated from the fixation process. Methods to repair FFPE DNA have been successful in human medicine for a variety of research and clinical applications, yet remain underutilized in veterinary medicine. Despite the available technology, our study is the first to evaluate the repair of FFPE derived DNA from veterinary species for single-nucleotide polymorphism (SNP) analysis using the Illumina OvineSNP50 BeadChip and Illumina FFPE QC and DNA Restore kit. To accomplish this, 48 ovine FFPE samples were run using the Illumina OvineSNP50 BeadChip with and without restoration. Compared to pre-restore data, we found increased sample call rates, SNP call frequency, and assay metrics for all samples post-restoration. Further, we utilized four sheep with available parallel fresh DNA and FFPE DNA to compare assay metrics and genotype calls between the two starting sample types. Although fresh samples generated increased call rates, we found 99% concordance in allele calls between restored FFPE and fresh DNA for all four samples. Our results indicate successful restoration and genotyping of ovine FFPE samples using this technology, with potential for utilization in other veterinary species.

Identifiants

pubmed: 35618986
doi: 10.1007/s11259-022-09937-0
pii: 10.1007/s11259-022-09937-0
pmc: PMC9873697
doi:

Substances chimiques

Formaldehyde 1HG84L3525
DNA 9007-49-2

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

131-139

Informations de copyright

© 2022. The Author(s).

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Auteurs

Amanda Kravitz (A)

Center for One Health Research, Department of Biomedical Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA.

Ron Tyler (R)

Center for One Health Research, Department of Biomedical Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA.

B Murali Manohar (BM)

Tamilnadu Veterinary and Animal Sciences University, Madhavaram Milk Colony, Chennai, 600051, Tamil Nadu, India.

B Samuel Masilamoni Ronald (BSM)

Tamilnadu Veterinary and Animal Sciences University, Madhavaram Milk Colony, Chennai, 600051, Tamil Nadu, India.

Michael T Collins (MT)

Department of Pathobiological Sciences, University of Wisconsin-Madison School of Veterinary Medicine, Madison, WI, USA.

Nammalwar Sriranganathan (N)

Center for One Health Research, Department of Biomedical Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA. nathans@vt.edu.

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Classifications MeSH