PCR protocol to identify an equine mutation associated with Warmblood Fragile Foal Syndrome(WFFS) v1

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 present 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. DOI dx.doi.org/10.17504/protocols.io.bw4fpgtn EXTERNAL LINK https://doi.org/10.1186/s13620-021-00206-1 PROTOCOL CITATION gerald.barry , Sharon Flanagan, Áine Rowe, Vivienne Duggan, Erin Markle, Maureen O’Brien 2021. PCR protocol to identify an equine mutation associated with Warmblood Fragile Foal Syndrome(WFFS). protocols.io https://dx.doi.org/10.17504/protocols.io.bw4fpgtn MANUSCRIPT CITATION please remember to cite the following publication along with this protocol Rowe LICENSE This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited CREATED Aug 03, 2021 LAST MODIFIED Aug 24, 2021 PROTOCOL INTEGER ID 52071 2 Citation: gerald.barry , Sharon Flanagan, Ã​Â​ine Rowe, Vivienne Duggan, Erin Markle, Maureen OâÂ​Â​Brien PCR protocol to identify an equine mutation associated with Warmblood Fragile Foal Syndrome(WFFS) https://dx.doi.org/10.17504/protocols.io.bw4fpgtn This is an open access protocol distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited