Identification of the causative mutation for inherited connective tissue disorders in equines and methods for testing for same

Abstract

Provided is a description of a mutation which is positively correlated with Warmblood Fragile Foal Syndrome Type 1 (WFFST1). The mutation is a G to A change at a specific location in the equine lysyl hydroxylase 1 (LH1) gene. Compositions and methods for use in diagnosing WFFST1 are provided.

We claim: 
     
         1 . A method for determining whether a horse is heterozygous, homozygous or has an absence of a Warmblood Fragile Foal Syndrome Type 1 (WFFST1) mutation, wherein the WFFST1 mutation comprises a change from G to A at position 147 in the sequence of SEQ ID NO:26, the method comprising: obtaining a biological sample from the horse and determining from the biological sample the presence or absence of the WFFST1 mutation, and identifying the horse as WFFST1 mutation homozygous by determining homozygosity for the WFFST1 mutation; or identifying the horse as WFFST1 mutation heterozygous by determining heterozygosity for the WFFST1 mutation; or identifying the horse as genetically normal with respect to WFFST1 by determining a homozygous absence of the WFFST1 mutation. 
     
     
         2 . The method of  claim 1 , wherein the determining the presence or absence of the WFFST1 mutation comprises isolating DNA or RNA from the biological sample and testing the isolated DNA and/or RNA for the presence or absence of the WFFST1 mutation. 
     
     
         3 . The method of  claim 1 , wherein the determining the presence or absence of the WFFST1 mutation comprises processing the biological sample to isolate DNA which comprises or might comprise the WFFST1 mutation, subsequently amplifying a segment of the isolated DNA which comprises or might comprise the WFFST1 mutation to obtain a DNA amplification product, and testing the DNA amplification product to determine the presence or absence of the WFFST1 mutation. 
     
     
         4 . The method of  claim 3 , wherein the determining the presence or absence of the WFFST1 mutation comprises separating RNA from the biological sample, creating a cDNA from the separated RNA, and testing the cDNA to determine the presence or absence of the WFFST1 mutation. 
     
     
         5 . The method of  claim 1 , wherein the determining the presence or absence of the WFFST1 mutation comprises isolating protein from the biological sample and testing the isolated protein for the presence of a protein comprising the sequence of SEQ ID NO:5, wherein the presence of a protein comprising the sequence of SEQ ID NO:5 establishes the presence of the WFFST1 mutation. 
     
     
         6 . A method for determining whether a genome of a horse comprises a Warmblood Fragile Foal Syndrome Type 1 (WFFST1) mutation, wherein the WFFST1 mutation comprises a change from G to A at position 4 in the sequence of SEQ ID NO:6, the method comprising: obtaining a biological sample from the horse and testing the biological sample to determine the presence or absence of the WFFST1 mutation. 
     
     
         7 . The method of  claim 6 , wherein the determining the presence of the mutation comprises determining the presence of SEQ ID NO:7. 
     
     
         8 . The method of  claim 6 , wherein the determining the absence of the mutation comprises determining the presence of SEQ ID NO:6. 
     
     
         9 . The method of  claim 6 , comprising determining the absence of the WFFST1 mutation and identifying the horse from which the biological sample was obtained as not affected with WFFST1. 
     
     
         10 . The method of  claim 6 , comprising determining the presence of the WFFST1 mutation and identifying the horse from which the biological sample was obtained as not genetically normal with respect to WFFST1. 
     
     
         11 . The method of  claim 6 , comprising determining homozygosity for the WFFST1 mutation and identifying the horse from which the biological sample was obtained as affected with or predisposed to WFFST1. 
     
     
         12 . The method of  claim 6 , comprising determining heterozygosity for the WFFST1 mutation and identifying the horse from which the biological sample was obtained as a carrier of the WFFST1 mutation. 
     
     
         13 . The method of  claim 6 , wherein the determining the presence of the WFFST1 mutation comprises detecting from the biological sample a polypeptide comprising the sequence of SEQ ID NO:5. 
     
     
         14 . A composition comprising an isolated nucleic acid, wherein the isolated nucleic acid comprises the sequence of SEQ ID NO:1.