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

The research discusses the development of a new, real-time PCR assay to detect the genetic mutation causing Warmblood Fragile Foal syndrome (WFFS) in a quick, reliable and cost-effective way without the need for sequencing.

Background on Warmblood Fragile Foal Syndrome

• Warmblood Fragile Foal syndrome (WFFS) is an inherited disorder that impacts the maturation of collagen in foals.
• This condition is usually fatal, with affected foals dying or having to be euthanised soon after birth.
• The syndrome is caused by a genetic mutation, specifically a single-point mutation at position 2032 of the equine PLOD1 gene, resulting in impairment of the normal enzyme.

Current Testing Method and Its Limitations

• A commercial test for identifying the causative genetic mutation of WFFS exists but it has some limitations.
• This test, licensed by Cornell University, involves amplifying a region of the PLOD1 gene that includes the mutation site, then sequencing that region and performing computational analysis.
• While this process is comprehensive, it is also costly and time-consuming, limiting the number of samples that can be analysed in a given timeframe.

The New Real-Time PCR Assay

• The researchers in this study have developed an alternative testing method: a real-time PCR assay.
• This PCR assay quickly and reliably distinguishes between the wild-type (original form) and WFFS-related mutations without the necessity for sequencing.
• The benefit of this real-time PCR approach is that it can analyse large numbers of samples efficiently and cost-effectively, increasing the throughput of testing.
• This means the new technique could speed up the detection process of the genetic mutation causing WFFS, thus contributing to early diagnosis, screening programs, and potential breeding strategies to reduce the prevalence of the syndrome.