Identification of canine leukocyte adhesion deficiency in dogs

• US 6,210,897 B1
• Filed: 09/10/1999
• Issued: 04/03/2001
• Est. Priority Date: 05/26/1999
• Status: Expired due to Term

• Current Assignee: Ake Hedhammar, Gunilla Wigh-Trowaldh, James Kijas, Leif Andersson, Sophie Gafvert
• Original Assignee: Ake Hedhammar, Gunilla Wigh-Trowaldh, James Kijas, Leif Andersson, Sophie Gafvert
• Inventors: Andersson, Leif, Kijas, James, Wigh-Trowaldh, Gunilla, Gafvert, Sophie, Hedhammar, Ake
• Primary Examiner(s): Sisson, Bradley L.
• Application Number:
  US09/393,554
• Time in Patent Office: 571 Days
• Field of Search: 435/6, 435/91.1, 435/91.2, 435/183, 536/23.1, 536/23.3, 536/24.3, 536/24.33, 536/25.3
• US Class Current: 435/6.13
• CPC Class Codes: C07K 14/70553 Integrin beta2-subunit-containing molecules, e.g. CD11, CD18
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/124 Animal traits, i.e. production traits, including athletic performance or the like
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers

Abstract

The present invention relates to an isolated nucleic acid molecule encoding canine leukocyte integrin β-2 subunit having a Cys36Ser missense mutation, wherein the missense mutation is indicative of a carrier of canine leukocyte adhesion deficiency. The present invention also relates to a method for identifying dogs which are carriers of or are affected with canine leukocyte adhesion deficiency. This method includes obtaining a biological sample from a dog and testing the biological sample for a Cys36Ser missense mutation in a gene encoding leukocyte integrin β-2 subunit, wherein the missense mutation in one allele is indicative of a carrier of canine leukocyte adhesion deficiency and the missense mutation in both alleles is indicative of a dog affected with canine leukocyte adhesion deficiency.

Claims

1. An isolated nucleic acid molecule encoding canine leukocyte integrin β

-2 subunit having a Cys36Ser missense mutation in the nucleic acid sequence according to SEQ. ID. No. 10, wherein said missense mutation is indicative of a carrier of canine leukocyte adhesion deficiency and said missense mutation is caused by a G→

C transversion at nucleotide 107 in the nucleic acid of SEQ. ID. No. 10.

2. An isolated nucleic acid molecule comprising:

a DNA molecule having the nucleotide sequence selected from the group consisting of SEQ. ID. No. 11, SEQ. ID. No. 12, SEQ. ID. No. 13, SEQ. ID. No. 14, SEQ. ID. No. 15, SEQ. ID. No. 16, SEQ. ID. No. 17, SEQ. ID. No. 18, SEQ. ID. No. 19, SEQ. ID. No. 20, SEQ. ID. No. 21, SEQ. ID. No. 22, and SEQ. ID. No. 23.

3. A method for identifying Irish Setter dogs which are carriers of or are affected with canine leukocyte adhesion deficiency, said method comprising:

obtaining a biological sample from an Irish Setter dog and

testing the biological sample for a Cys36Ser missense mutation in a gene encoding leukocyte integrin β

-2 subunit, wherein the Cvs36Ser missense mutation is caused by a G→

C transversion at nucleotide 107 in the nucleic acid of SEQ. ID. No. 10 and the missense mutation in one allele is indicative of a carrier of canine leukocyte adhesion deficiency and the missense mutation in both alleles is indicative of an Irish Setter dog affected with canine leukocyte adhesion deficiency.

4. The method according to claim 3, wherein said testing is carried out by an oligonlicleotide ligation assay.

5. The method according to claim 3, wherein said testing is carried out by direct sequence analysis.

6. The method according to claim 3, wherein said detecting is carried out by subjecting the biological sample to conditions effective to hybridize DNA molecules with the missense mutation to a probe specific for the sequence of the missense mutation.

7. The method according to claim 3, wherein said detecting is carried out by digesting the biological sample with a restriction endonuclease.

8. The method according to claim 3, wherein said testing is carried out by ligase detection reaction.

9. The method according to claim 3, wherein said testing is carried out by single strand polymorphism assay.

10. The method according to claim 3, wherein said testing is carried out by ligase chain reaction.

11. The method according to claim 3, wherein said testing is carried out by PCR-based restriction fragment length polymorphism.

12. The method according to claim 3, wherein said testing comprises:

amplifying a region of the gene encoding a leukocyte integrin β

-2 subunit to provide an amplified fragment before detecting any Cvs36Ser missense mutation present in the biological sample.

13. The method according to claim 3, wherein the nucleic acid is a deoxyribonucleic acid.

14. The method according to claim 3, Wherein the nucleic acid is a messenger ribonucleic acid.

15. The method according to claim 3, wherein the biological sample is any tissue containing genomic DNA.

16. The method according to claim 15, wherein the biological sample is blood, hair, check scrapings, semen, tissue biopsy, or saliva.