Composition and methods for detection of von willebrand's disease

An assay kit for screening for a canine von Willebrand's Factor gene comprising:

• a) an oligonucleotide comprising at least 10 contiguous nucleotides of the nucleic acid sequence of SEQ ID NO:1, and capable of hybridizing with the nucleotide sequence encoding canine von Willebrand's Factor,
• b) reagents for hybridization of the oligonucleotide to a complementary nucleic acid sequence;
• and
• c) container means for a) and b).

An assay kit for screening for a canine von Willebrand's Factor gene comprising:

• a) an oligonucleotide comprising at least 10 contiguous nucleotides of the nucleic acid sequence of SEQ ID NO:2, and capable of hybridizing with the nucleotide sequence encoding canine von Willebrand's Factor;
• b) reagents for hybridization of the oligonucleotide to a complementary nucleic acid sequence;
• and
• c) container means for a) and b).

An assay kit for screening for a canine von Willebrand's Factor gene comprising:

• a) an oligonucleotide comprising contiguous nucleotides of the nucleic acid sequence that is complementary to the sequence of SEQ ID NO:1 having a mutation at nucleotide 172, and capable of specifically hybridizing to the complementary nucleotide sequence;
• b) reagents for hybridization of the oligonucleotide to a complementary nucleic acid sequence;
• and
• c) container means for a) and b).

An assay kit for screening for a canine von Willebrand's Factor gene comprising:

• a) an oligonucleotide comprising contiguous nucleotides of the nucleic acid sequence that is complementary to the sequence of SEQ ID NO:2 having a mutation at nucleotide 384, and capable of specifically hybridizing to the complementary nucleotide sequence;
• b) reagents for hybridization of the oligonucleotide to a complementary nucleic acid sequence;
• and
• c) container means for a) and b).

A method for detecting a mutated canine von Willebrand's Factor gene in a canine DNA sample comprising the steps of:

• a) amplifying the DNA sample by polymerase chain reaction to produce polymerase chain reaction products, wherein the polymerase chain reaction uses primers that produce a restriction site in a normal allele but not in a mutant allele, wherein the mutation in the mutant allele is a substitution at nucleotide 172 of the nucleotide sequence encoding canine von Willebrand's Factor polypeptide, wherein the nucleotide sequence is capable of hybridizing under high stringency conditions to the complementary sequence of the sequence of SEQ ID NO:1;
• b) digesting the polymerase chain reaction products with a restriction enzyme specific to the restriction site of the primers to produce DNA fragments;
• and
• c) detecting the DNA fragments, thereby detecting a mutated canine von Willebrand's Factor gene.

A method for detecting a mutated canine von Willebrand's Factor gene in a canine DNA sample comprising the steps of:

• a) amplifying the DNA sample by polymerase chain reaction to produce polymerase chain reaction products, wherein the polymerase chain reaction uses primers that are complementary to sequences of the introns flanking the exon, wherein the exon of the mutant allele has a deletion at nucleotide 384 of the nucleotide sequence encoding canine von Willebrand's Factor polypeptide, wherein the nucleotide sequence is capable of hybridizing under high stringency conditions to the complementary sequence of the sequence of SEQ ID NO:2;
• b) digesting the polymerase chain reaction products with a restriction enzyme specific to the restriction site of the mutant allele to produce DNA fragments;
• and
• c) detecting the DNA fragments, thereby detecting a mutated canine von Willebrand's Factor gene.