Journal
LIFE-BASEL
Volume 12, Issue 6, Pages -Publisher
MDPI
DOI: 10.3390/life12060851
Keywords
CMTs; FFPE; validation; repeatability; dog tumors
Categories
Funding
- Italian Ministry of Health [IZS AM 06/19 RC]
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Canine mammary tumors share characteristics with human breast cancers, and the NGS panel developed in this study is a useful tool for expanding BRCA gene knowledge in the veterinary field and for comparative oncology research.
Mammary carcinomas are the most common neoplasms observed in women and in female dogs. Canine mammary tumors show epidemiological, clinical, genetic, and prognostic characteristics comparable to human breast cancers. The recent introduction of next generation sequencing (NGS) technologies has greatly improved research and diagnostics for humans, while these new tools still need to be implemented in animal models. In this study we developed and validated an AmpliSeq Panel assay for the identification of BRCA variants in twenty-two different dogs. The amplicon mean coverage was 5499x and uniformity was higher than 98% in all samples. The results of germline single nucleotide variants (SNVs) and insertions/deletions (INDELs) were fully concordant regardless of the types of samples considered (blood, fresh and FFPE tissues). Moreover, despite the high DNA degradation observed in older FFPE blocks (>5 years), the assay allowed full coverage of all amplicons for downstream analyses. We consider the NGS panel developed in this study as a useful tool for expanding information on BRCA genes in the veterinary field and for human health from a comparative oncology perspective.
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