Diagnostic assay for tissue transplantation status
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.
1 . A method, comprising:
(a) extracting DNA comprising donor and recipient DNA fragments from a substantially cell-free sample of blood plasma, blood serum or urine of an organ transplant recipient;
(b) amplifying the DNA extracted in (a) via an amplification reaction which selectively amplifies DNA comprising copy number deletion (CND) polymorphism(s) which indicate the DNA is donor-derived, wherein the amplified DNA after (b) comprises a detectable level of extracellular circulatory donor DNA fragments which comprise the CND polymorphism(s); and
(c) quantifying the level of DNA comprising the CND polymorphism(s) which indicate the DNA is donor derived based on the results of the amplification reaction, wherein the CND(s) comprises at least one copy number variant polymorphism selected from the group consisting of CNP ID: CNVR358.1, CNVR217.1, CNVR483.1 CNVR451_full, CNVR376.1, CNVR381.1, 88, CNVR431.1, 262, CNVR1138.2, CNVR1138.3, CNVR966.1, CNVR1037.1, CNVR952.1, CNVR801_full, CNVR842.1, CNVR1041_full, CNVR1058.1, CNVR894.2, CNVR1648.2, CNVR1438_full, CNVR1576.1, CNVR1419.1, CNVR1685.1, CNVR1610.1, CNVR1341.1, CNVR1464.1, CNVR2196.1, CNVR1894.1, CNVR2221_full, CNVR1937.1, CNVR1935_full, CNVR1819.6, CNVR2172 full, CNVR2168 full, 726, CNVR2613_full, CNVR2535_full, CNVR2344_full, CNVR2469.1, CNVR2304.1, 896, CNVR2939.3, CNVR2939.2, CNVR2799.1, CNVR3004.1, CNVR2906.1, CNVR2972 full, CNVR2859 full, 933, CNVR3009.1, CNVR3472 full, CNVR3319.1, CNVR3495.1, 1103, CNVR3451.1, CNVR3609.1, CNVR3753_full, CNVR3935.1, CNVR4014.1, CNVR3831.1, CNVR3689.1, CNVR4250.1, CNVR4331.1, CNVR4374.1, CNVR4203.1, CNVR4332.1, CNVR4841.1, CNVR4665.1, CNVR4886.1, CNVR4596.1, CNVR4906.1, 1730, CNVR5122.1, CNVR5294.1, CNVR5429.1, CNVR5853 full, CNVR5923.1, CNVR5850.1, CNVR5871.1, CNVR6133.1, CNVR6211.1, CNVR6084.1, CNVR6074 full, CNVR6357.1, CNVR6540.1, CNVR6670.1, CNVR6676.1, CNVR6782.1, CNVR7144.1, CNVR7096.1, CNVR7301.1, CNVR7344.1, CNVR7543.1, CNVR7581 full, 2434, 2454, CNVR7808.1, CNVR7849.1, CNVR7796 full, 2559, CNVR8147.1, CNVR8114.4, and CNVR8154.1.
2 . The method of claim 1 , wherein the level of DNA comprising the CND polymorphism(s) which indicate the DNA is donor derived produced in (b) are not determined by nucleotide sequencing analysis or bi-allelic microchimerism analysis.
3 . The method of claim 1 , wherein the CND is CNVR376.1.
4 . The method of claim 1 , wherein the organ transplant recipient received an organ selected from the group consisting of a kidney, heart, lung, pancreatic islet, liver, intestine and skin.
5 . The method of claim 1 , wherein the organ transplant recipient received a kidney.
6 . The method of claim 1 , wherein the amplification reaction is digital PCR.