Non-invasive diagnosis of graft rejection in organ transplant patients

  • US 10,494,669 B2
  • Filed: 08/23/2018
  • Issued: 12/03/2019
  • Est. Priority Date: 11/06/2009
  • Status: Active Grant
First Claim
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1. A method of detecting donor-specific circulating cell-free nucleic acids in a heart transplant recipient, the method comprising:

  • (a) genotyping a heart transplant donor to obtain a single nucleotide polymorphism (SNP) profile of said heart transplant donor;

    (b) genotyping a heart transplant recipient to obtain a SNP profile of the heart transplant recipient;

    (c) obtaining a biological sample from said heart transplant recipient after said heart transplant recipient has received a heart transplant from said heart transplant donor, wherein said biological sample is selected from the group consisting of blood, serum and plasma, and wherein said biological sample comprises circulating cell-free nucleic acids from said heart transplant;

    (d) performing one or more quantitative polymerase chain reaction (PCR) reactions on said circulating cell-free deoxyribonucleic acid obtained from said biological sample from said heart transplant recipient, wherein said one or more quantitative PCR reactions amplify at least 10 different deoxyribonucleic acid targets, and wherein said at least 10 different deoxyribonucleic acid targets comprise at least 10 informative SNPs; and

    (e) quantifying an amount of heart transplant-derived circulating cell-free deoxyribonucleic acid from said heart transplant using markers distinguishable between said heart transplant recipient and said heart transplant donor, wherein said markers distinguishable between said heart transplant recipient and said heart transplant donor are said at least 10 informative SNPs, wherein said quantified amount of heart transplant-derived circulating cell-free deoxyribonucleic acid from said heart transplant using markers distinguishable between said heart transplant recipient and said donor comprises at least 0.03% of the total circulating cell free deoxyribonucleic acid from said biological sample from said heart transplant recipient.

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