Methods for detection of donor-derived cell-free dna
AssigneeNatera, Inc.
InventorsSolomon MOSHKEVICH, Bernhard ZIMMERMANN, Tudor Pompiliu CONSTANTIN, Huseyin Eser KIRKIZLAR, Allison RYAN, Styrmir SIGURJONSSON, Felipe ACOSTA ARCHILA, Ryan SWENERTON
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Abstract
The present disclosure provides methods for determining the status of an allograft within a transplant recipient from genotypic data measured from a mixed sample of DNA comprising DNA from both the transplant recipient and from the donor. The mixed sample of DNA may be preferentially enriched at a plurality of polymorphic loci in a way that minimizes the allelic bias, for example using massively multiplexed targeted PCR.
Claims22
A method of preparing a blood sample of a transplant recipient, comprising:
- a) extracting DNA from the blood sample of the transplant recipient, wherein the DNA comprises donor-derived cell-free DNA and recipient-derived cell-free DNA;
- b) performing targeted amplification at 200-50,000 target loci in a single reaction volume using 200-50,000 primer pairs to obtain amplification products, wherein the target loci comprise polymorphic loci and non-polymorphic loci;
- c) receiving data corresponding to at least one of donor height, donor weight, or donor transplant mass;
- and
- d) determining, based at least in part on the received data, a cutoff threshold value for the blood sample of the transplant recipient, the cutoff threshold value is indicative of an amount of the donor-derived cell-free DNA in the blood sample.
Dependencies are inferred from the claim text.