Diagnostic test for Collie Eye Anomaly

• US 7,723,507 B2
• Filed: 09/22/2008
• Issued: 05/25/2010
• Est. Priority Date: 10/20/2004
• Status: Active Grant

• Current Assignee: Cornell Research Foundation Incorporated (Cornell University)
• Original Assignee: Cornell Research Foundation Incorporated (Cornell University)
• Inventors: Aguirre, Gustavo D., Akey, Dayna, Kukekova, Anna V., Ostrander, Elaine, Acland, Gregory M., Goldstein, Orly
• Primary Examiner(s): Myers; Carla
• Application Number:
  US12/234,803
• Publication Number:
  US 20090176225A1
• Time in Patent Office: 610 Days
• Field of Search: None
• US Class Current: 536/24.33
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 1/6888 for detection or identification of organisms
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: C12Q 2600/16 Primer sets for multiplex assays

Abstract

The invention relates to a method for identifying dogs which are genetically normal, heterozygous for, or homozygous for the mutation primarily responsible for Collie eye anomaly (CEA). The method comprises the steps of obtaining a biological sample from a dog and testing DNA in the biological sample for the presence or absence of a 7.8 kilobase deletion within chromosome 37 in which the CEA mutation is located. No deletion is indicative of a normal dog. A deletion on one allele of chromosome 37 is indicative of a dog that is heterozygous for the CEA mutation. A deletion in both alleles of chromosome 37 are indicative of a dog that is homozygous for the CEA mutation. Also provided is a kit for identifying a dog as normal, heterozygous for, or homozygous for the CEA mutation.

Claims

1. A kit for identifying a dog as normal, heterozygous for, or homozygous for a Collie Eye Anomaly mutation in a region of chromosome 37, wherein the mutation consists of a deletion of nucleotides 9,302 to 17,101 of SEQ ID NO:

1, comprising;

a set of primers, wherein the only primers in the kit are a first primer, a second primer, a third primer and a fourth primer, wherein the first primer, the second primer, the third primer, and the fourth primer each consist of a fragment of SEQ ID NO;

1 or the complement thereof, wherein the first primer binds to a sequence upstream of position 9,301 of SEQ ID NO;

1, the fourth primer binds to a sequence downstream of position 17,102 of SEQ ID NO;

1 and second and third primers binds to a sequence between nucleotides 9,302 and 17,101 of SEQ ID NO;

1, wherein amplification of an allele that does not comprise the mutation with the first and the second primers produces a first amplification product that comprises the nucleotide 9,302 of SEQ ID NO;

1, amplification of an allele that does not comprise the mutation using the third and fourth primers produces a second amplification product that comprises the nucleotide 17,101 of SEQ ID NO;

1, and amplification using the first and the fourth primer produces a third amplification product, wherein the third amplification product comprises SEQ ID NO;

14 or a fragment thereof wherein the fragment thereof comprises contiguous nucleotides 89 and 90 of SEQ ID NO;

14.

2. The kit of claim 1, wherein the first primer has the sequence of SEQ ID NO:

2, the second primer has the sequence of SEQ ID NO;

4, the third primer has the sequence of SEQ ID NO;

6 and the fourth primer has the sequence of SEQ ID NO;

8.