公开(公告)号：US20240229136A9

公开(公告)日：2024-07-11

申请号：US18279006

申请日：2022-02-24

Applicant: Natera, Inc.

Inventor： Zachary DEMKO ,  Ebad AHMED ,  Ryan SWENERTON ,  Paul VAN HUMMELEN

IPC: C12Q1/6883 ,  C12Q1/6869

CPC classification number: C12Q1/6883 ,  C12Q1/6869 ,  C12Q2600/156

Abstract: The present disclosure provides methods of amplifying and sequencing DNA, comprising: extracting cell-free DNA from a blood, plasma, serum or urine sample of a transplant recipient who has received transplantation of one or more organs including simultaneous or sequential transplantation of multiple organs, wherein the extracted cell-free DNA comprises donor-derived cell-free DNA and recipient-derived cell-free DNA; performing targeted amplification at 200-50,000 target loci in a single reaction volume using 200-50,000 primer pairs, wherein the target loci comprise polymorphic loci and non-polymorphic loci; sequencing the amplification products by high-throughput sequencing to obtain a sequencing reads and quantifying the amount of donor-derived cell-free DNA and the amount of total cell-free DNA based on the sequencing reads; and determining whether the amount of donor-derived cell-free DNA or a function thereof exceeds a cutoff threshold indicating transplant rejection or graft injury.

公开(公告)号：US20240132960A1

公开(公告)日：2024-04-25

申请号：US18279006

申请日：2022-02-24

Applicant: Natera, Inc.

Inventor： Zachary DEMKO ,  Ebad AHMED ,  Ryan SWENERTON ,  Paul VAN HUMMELEN

IPC: C12Q1/6883 ,  C12Q1/6869

CPC classification number: C12Q1/6883 ,  C12Q1/6869 ,  C12Q2600/156

Abstract: The present disclosure provides methods of amplifying and sequencing DNA, comprising: extracting cell-free DNA from a blood, plasma, serum or urine sample of a transplant recipient who has received transplantation of one or more organs including simultaneous or sequential transplantation of multiple organs, wherein the extracted cell-free DNA comprises donor-derived cell-free DNA and recipient-derived cell-free DNA; performing targeted amplification at 200-50,000 target loci in a single reaction volume using 200-50,000 primer pairs, wherein the target loci comprise polymorphic loci and non-polymorphic loci; sequencing the amplification products by high-throughput sequencing to obtain a sequencing reads and quantifying the amount of donor-derived cell-free DNA and the amount of total cell-free DNA based on the sequencing reads; and determining whether the amount of donor-derived cell-free DNA or a function thereof exceeds a cutoff threshold indicating transplant rejection or graft injury.

公开(公告)号：US20230203573A1

公开(公告)日：2023-06-29

申请号：US17925693

申请日：2021-05-27

Applicant: Natera, Inc.

Inventor： Ryan SWENERTON ,  Bernhard ZIMMERMANN ,  Ebad AHMED ,  Nathan LIANG ,  Allison RYAN ,  Fei LU ,  Paul VAN HUMMELEN

IPC: C12Q1/6851 ,  C12Q1/686 ,  C12Q1/6876

CPC classification number: C12Q1/6851 ,  C12Q1/686 ,  C12Q1/6876 ,  C12Q2600/158

Abstract: The present disclosure provides methods for quantifying the amount of total cell-free DNA in a biological sample, comprising: isolating cell-free DNA from the biological sample, wherein a first Tracer DNA composition is added before or after isolation of the cell-free DNA; performing targeted amplification at 100 or more different target loci in a single reaction volume using 100 or more different primer pairs; sequencing the amplification products by high-throughput sequencing to generate sequencing reads; and quantifying the amount of total cell-free DNA using sequencing reads derived from the first Tracer DNA composition.