Chromosomal assessment to differentiate histiocytic malignancy from lymphoma and hemangiosarcoma in dogs

Abstract

This invention relates generally to the discovery of an improved method to differentiate histiocytic malignancy from lymphoma or hemangiosarcoma in dogs.

What is claimed is: 
     
         1 . A kit for differentiating a histiocytic malignancy from a lymphoma or an hemangiosarcoma in a dog, the kit comprising at least three nucleic acid probes, primers, or pair of primers, wherein a first of the at least three nucleic acid probes, primers, or pair of primers is capable of specifically detecting CFA 2 in a biological sample comprising neoplastic cells obtained from a dog, a second of the at least three nucleic acid probes, primers, or pair of primers is capable of specifically detecting CFA 16 in the biological sample, and a third of the at least three nucleic acid probes, primers, or pair of primers is capable of specifically detecting CFA 31 in the biological sample. 
     
     
         2 . The kit of  claim 1 , wherein the biological sample is selected from the group consisting of a fine needle aspirate and a biopsy. 
     
     
         3 . The kit of  claim 1 , wherein the biological sample is a cytological smear, a fresh-frozen sample, a fresh sample, a fixed sample, and/or a paraffin-embedded sample. 
     
     
         4 . The kit of  claim 1 , wherein the kit further comprises one or more additional components selected from the group consisting of blocking agents, detectable labels or labeling agents, and reagents for hybridization of the at least three nucleic acid probes, primers, or pair of primers to CFA 2, CFA 16, and CFA 31 in the biological sample. 
     
     
         5 . The kit of  claim 1 , wherein the at least one nucleic acid probe, primer, or pair of primers is appropriate for use in measuring copy number by a technique selected from the group consisting of fluorescence in situ hybridization (FISH); polymerase chain reaction (PCR); comparative genomic hybridization (CGH); next generation sequencing, and/or droplet digital PCR (ddPCR). 
     
     
         6 . The kit of  claim 5 , wherein the kit comprises at least one pair of primers that can be employed to measure copy number of CFA 2, CFA 16, and/or CFA 31 in the biological sample. 
     
     
         7 . The kit of  claim 1 , wherein one or more of the at least three nucleic acid probes, primers, or pair of primers is detectably labeled. 
     
     
         8 . The kit of  claim 7 , wherein the first nucleic acid probe, primer, or pair of primers, the second nucleic acid probe, primer, or pair of primers, and the third nucleic acid probe, primer, or pair of primers are labeled with different detectable labels. 
     
     
         9 . The kit of  claim 8 , wherein the first, second, and third nucleic acid probes, primers, or pairs of primers are labeled with different fluorophores. 
     
     
         10 . The kit of  claim 1 , wherein the kit further comprises a nuclear stain, optionally wherein the nuclear stain is 4′,6-diamidino-2-phenylindole (DAPI). 
     
     
         11 . The kit of  claim 1 , further comprising at least one additional probe, primer, or pair of primers capable of specifically detecting a dog chromosome other than CFA 2, CFA 16, and/or CFA 31 and for measuring a copy number of the dog chromosome other than CFA 2, CFA 16, and/or CFA 31 to serve as a copy number control. 
     
     
         12 . The kit of  claim 11 , wherein the kit comprises a first nucleic acid probe, primer, or pair of primers that is capable of specifically detecting copy number at CFA 2; a second nucleic acid probe, primer, or pair of primers that is capable of specifically detecting copy number at CFA 16; a third nucleic acid probe, primer, or pair of primers that is capable of specifically detecting copy number at CFA 31; and at least one additional probe, primer, or pair of primers capable of specifically detecting a dog chromosome other than CFA 2, CFA 16, or CFA 31; and further wherein the first, second, third, and at least one additional nucleic acid probes, primers, or pairs of primers are labeled with different detectable labels. 
     
     
         13 . The kit of  claim 11 , wherein the differently labeled first, second, third, and at least one additional nucleic acid probes, primers, or pairs of primers are suitable for use in a three or four color cytogenetic assay. 
     
     
         14 . The kit of  claim 10 , wherein the first of the at least three nucleic acid probes, primers, or pair of primers is suitable for measuring a copy number of chr2:31,017,808-31,586,959 of canfam2; the second of the at least three nucleic acid probes, primers, or pair of primers is suitable for measuring a copy number of chr16:31,674,146-32,429,051 of canfam2; and the third of the at least three nucleic acid probes, primers, or pair of primers is suitable for measuring a copy number of chr31:33,346,317-33,709,291 of canfam2. 
     
     
         15 . The kit of  claim 1 , wherein the kit further comprises instructions for use of the kit in measuring copy numbers of CFA 2, CFA 16, and CFA 31, and optionally the dog chromosome other than CFA 2, CFA 16, and/or CFA 31 in the biological sample.