Open AccessDevelopment of a real-time PCR assay to detect the single nucleotide polymorphism causing Warmblood Fragile Foal Syndrome
Author(s) -
Sharon Flanagan,
Áine Rowe,
Vivienne Duggan,
Erin Markle,
Megan B. O’Brien,
Gerald Barry
Publication year - 2021
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0259316
Subject(s) - foal , warmblood , sanger sequencing , biology , genetics , single nucleotide polymorphism , gene , microbiology and biotechnology , mutation , genotype , horse , paleontology
Warmblood Fragile Foal syndrome (WFFS) is an autosomal recessive condition that affects the maturation of collagen in affected foals. Foals affected with the disease typically die or are euthanised shortly after birth. WFFS is caused by a single nucleotide change at position 2032 of the equine PLOD1 gene, causing an impairment of the wild-type enzyme. A commercial test for the causative genetic mutation is currently available from companies operating under licence from Cornell University but it has limitations. This test requires amplification of a region of the PLOD1 gene encompassing the site of interest, followed by Sanger sequencing of that region and computational analysis. We describe here the development of an alternative, real-time PCR based assay that rapidly and reliably differentiates between the wild-type and WFFS associated nucleotides without the need for sequencing, thus increasing the potential for high throughput analysis of large numbers of samples in a cost-effective manner.