Diagnostic assay for tissue transplantation status

Abstract

A method of detecting circulatory nucleic acids from a transplanted tissue in a transplant recipient, the method comprising: amplifying circulatory nucleic acids from a transplanted tissue in a blood sample from the transplant recipient, and detecting amplification of a copy number deletion (CND) polymorphism from the transplanted tissue, and related methods for determining the status of a donor tissue transplanted into a recipient.

What is claimed is: 
     
         1 . A method of detecting circulatory nucleic acids from a transplanted tissue in a transplant recipient, the method comprising:
 amplifying circulatory nucleic acids from a transplanted tissue in a blood sample from the transplant recipient, and   detecting amplification of a copy number deletion (CND) polymorphism from the transplanted tissue, wherein the CND polymorphism is selected from the group consisting of:
 chromosome 1, nucleotides 157,134,152-157,136,674, 
 chromosome 18, nucleotides 53,097,735-53,099,702, 
 chromosome 8, nucleotides 112,363,260-112,365,469, 
 chromosome 14, nucleotides 21,951,540-21,952,070, 
 chromosome 16, nucleotides 25,247,611-25,250,595, 
 chromosome 7, nucleotides 24,004,755-24,006,584, 
 chromosome 13, nucleotides 37,955,318-37,958,191, 
 chromosome 8, nucleotides 4,110,308-4,112,335, 
 chromosome 18, nucleotides 33,560,073-33,560,645, and 
 chromosome 10, nucleotides 108,020,303-108,022,518. 
   
     
     
         2 . The method of  claim 1  wherein the transplanted tissue is an organ selected from the group consisting of a kidney, heart, lung, pancreatic islet, liver, intestine and skin. 
     
     
         3 . The method of  claim 1  wherein the transplanted tissue is a microtissue or stem cells. 
     
     
         4 . The method of  claim 1  wherein the transplanted tissue is a kidney. 
     
     
         5 . The method of  claim 1 , wherein circulatory nucleic acids are amplified using digital PCR. 
     
     
         6 . The method of  claim 1 , wherein at least 2 CND polymorphisms from the transplanted tissue are detected. 
     
     
         7 . The method of  claim 1 , wherein at least 3 CND polymorphisms from the transplanted tissue are detected. 
     
     
         8 . The method of  claim 1 , wherein the selected CND polymorphism is nullisomic with regard to circulatory nucleic acids that are not from the transplanted tissue. 
     
     
         9 . A method for determining the status of a donor tissue transplanted 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 deletion (CND) 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. 
     
     
         10 . The method of  claim 9 , wherein the tissue transplanted is a kidney. 
     
     
         11 . A circulatory nucleic acid having a copy number deletion (CND) polymorphism, wherein the CND polymorphism is selected from the group consisting of:
 chromosome 1, nucleotides 157,134,152-157,136,674,   chromosome 18, nucleotides 53,097,735-53,099,702,   chromosome 8, nucleotides 112,363,260-112,365,469,   chromosome 14, nucleotides 21,951,540-21,952,070,   chromosome 16, nucleotides 25,247,611-25,250,595,   chromosome 7, nucleotides 24,004,755-24,006,584,   chromosome 13, nucleotides 37,955,318-37,958,191,   chromosome 8, nucleotides 4,110,308-4,112,335,   chromosome 18, nucleotides 33,560,073-33,560,645, and   chromosome 10, nucleotides 108,020,303-108,022,518.