Diagnostic assay for tissue transplantation status

Abstract

The present disclosure relates generally to tissue, including organ, transplantation. Taught herein is a genetic-based diagnostic assay to ascertain the status of a tissue transplantation procedure in a recipient based on a characterization of circulating or other cell free nucleic acid material. The instant disclosure enables kits, primers and protocols for ascertaining the status of a transplantation procedure.

1 . A method for determining the status of a donor tissue implanted into a recipient, said method comprising screening a sample from the recipient for the presence or absence of circulatory nucleic acids which have a copy number variant (CNV) polymorphism which indicates they are donor-derived nucleic acids wherein the presence of donor nucleic acids is indicative of cellular damage of the transplanted tissue and the absence of donor nucleic acids or a level of donor nucleic acids relative to a control is indicative of no cellular damage or cellular damage to an acceptable level. 
     
     
         2 . The method of  claim 1  wherein the nucleic acids in the sample are tested for a CNV from a panel of CNV sites wherein a given recipient or donor is characterized by being nullisomic for at least one of the CNV sites and the other of the recipient or donor is not. 
     
     
         3 . The method of  claim 1  wherein the presence of the CNV is not determined by nucleotide sequence analysis or bi-allelic microchimerism analysis. 
     
     
         4 . The method of  claim 2  wherein the CNV is a copy number deletion (CND) polymorphism. 
     
     
         5 . The method of  claim 4  wherein the CND site is selected from the list set forth in Table 2 wherein the recipient is characterized by having a null genotype for at least one CND site. 
     
     
         6 . The method of  claim 1  wherein the tissue transplanted is an organ selected from the group consisting of a kidney, heart, lung, pancreatic islet, liver, intestine and skin. 
     
     
         7 . The method of  claim 1  wherein the tissue transplanted is a limb selected from the group consisting of a leg, arm, hand and foot. 
     
     
         8 . The method of  claim 1  wherein the tissue transplanted is an appendage selected from the group consisting of a toe, nose and ear. 
     
     
         9 . The method of  claim 1  wherein the tissue is microtissue or stem cells. 
     
     
         10 . The method of  claim 1  wherein the tissue transplanted is a kidney. 
     
     
         11 . The method of  claim 1  wherein the sample is selected from the group consisting of plasma, whole blood, serum, urine, pus, respiratory fluid, lymph fluid, feces, bile, saliva, sputum, semen, vaginal flow, cerebrospinal fluid, brain fluid, ascites, milk, secretions from the genitourinary tract and a lavage of a tissue or organ. 
     
     
         12 . The method of  claim 11  wherein the sample is plasma, whole blood or serum. 
     
     
         13 . The method of  claim 11  wherein the sample is urine. 
     
     
         14 . A transplantation protocol for transplanting tissue from a donor to a recipient, said protocol comprising transplanting the tissue and then monitoring a sample from the recipient for the presence of nucleic acids which are not the recipients wherein if donor nucleic acids are detected, the recipient is subjected to immunosuppression therapy or if the patient is on immunosuppression therapy, the therapy is altered. 
     
     
         15 . The method of  claim 14  wherein the nucleic acids in the sample are tested for a CNV from a panel of CNV sites wherein a given recipient or donor is characterized by being nullisomic for at least one of the CNV sites and the other of the recipient or donor is not. 
     
     
         16 . The method of  claim 14  wherein the presence of the CNV is not determined by nucleotide sequence analysis or bi-allelic microchimerism analysis. 
     
     
         17 . The method of  claim 15  wherein the CNV is a copy number deletion (CND) polymorphism. 
     
     
         18 . The method of  claim 15  wherein the CND site is selected from the list set forth in Table 2 wherein the recipient is characterized by having a null genotype for at least one CND site. 
     
     
         19 . The method of  claim 14  wherein the tissue transplanted is an organ selected from the group consisting of a kidney, heart, lung, pancreatic islet, liver, intestine and skin. 
     
     
         20 . The method of  claim 14  wherein the tissue transplanted is a limb selected from the group consisting of a leg, arm, hand and foot. 
     
     
         21 . The method of  claim 14  wherein the tissue transplanted is an appendage selected from the group consisting of a toe, nose and ear. 
     
     
         22 . The method of  claim 14  wherein the tissue is microtissue or stem cells. 
     
     
         23 . The method of  claim 14  wherein the tissue transplanted is a kidney. 
     
     
         24 . The method of  claim 14  wherein the sample is selected from the group consisting of plasma, whole blood, serum, urine, pus, respiratory fluid, lymph fluid, feces, bile, saliva, sputum, semen, vaginal flow, cerebrospinal fluid, brain fluid, ascites, milk, secretions from the genitourinary tract and a lavage of a tissue or organ. 
     
     
         25 . The method of  claim 23  wherein the sample is plasma, whole blood or serum. 
     
     
         26 . The method of  claim 24  wherein the sample is urine. 
     
     
         27 .- 29 . (canceled)