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Development of a real-time PCR assay to detect the single nucleotide polymorphism causing Warmblood Fragile Foal Syndrome

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...

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Autores principales: Flanagan, Sharon, Rowe, Áine, Duggan, Vivienne, Markle, Erin, O’Brien, Maureen, Barry, Gerald
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8575260/
https://www.ncbi.nlm.nih.gov/pubmed/34748589
http://dx.doi.org/10.1371/journal.pone.0259316
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author Flanagan, Sharon
Rowe, Áine
Duggan, Vivienne
Markle, Erin
O’Brien, Maureen
Barry, Gerald
author_facet Flanagan, Sharon
Rowe, Áine
Duggan, Vivienne
Markle, Erin
O’Brien, Maureen
Barry, Gerald
author_sort Flanagan, Sharon
collection PubMed
description 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.
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spelling pubmed-85752602021-11-09 Development of a real-time PCR assay to detect the single nucleotide polymorphism causing Warmblood Fragile Foal Syndrome Flanagan, Sharon Rowe, Áine Duggan, Vivienne Markle, Erin O’Brien, Maureen Barry, Gerald PLoS One Lab Protocol 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. Public Library of Science 2021-11-08 /pmc/articles/PMC8575260/ /pubmed/34748589 http://dx.doi.org/10.1371/journal.pone.0259316 Text en © 2021 Flanagan et al https://creativecommons.org/licenses/by/4.0/This is an open access article 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.
spellingShingle Lab Protocol
Flanagan, Sharon
Rowe, Áine
Duggan, Vivienne
Markle, Erin
O’Brien, Maureen
Barry, Gerald
Development of a real-time PCR assay to detect the single nucleotide polymorphism causing Warmblood Fragile Foal Syndrome
title Development of a real-time PCR assay to detect the single nucleotide polymorphism causing Warmblood Fragile Foal Syndrome
title_full Development of a real-time PCR assay to detect the single nucleotide polymorphism causing Warmblood Fragile Foal Syndrome
title_fullStr Development of a real-time PCR assay to detect the single nucleotide polymorphism causing Warmblood Fragile Foal Syndrome
title_full_unstemmed Development of a real-time PCR assay to detect the single nucleotide polymorphism causing Warmblood Fragile Foal Syndrome
title_short Development of a real-time PCR assay to detect the single nucleotide polymorphism causing Warmblood Fragile Foal Syndrome
title_sort development of a real-time pcr assay to detect the single nucleotide polymorphism causing warmblood fragile foal syndrome
topic Lab Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8575260/
https://www.ncbi.nlm.nih.gov/pubmed/34748589
http://dx.doi.org/10.1371/journal.pone.0259316
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