Method and kit for detecting presence or proportion of donors in acceptor sample

A sample and donor technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of unsuitability for regular monitoring, low detection sensitivity, high detection cost, and achieve intuitive quantitative results , The effect of low detection sensitivity and short detection period

Pending Publication Date: 2022-06-21
XIAMEN UNIV
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Problems solved by technology

However, the above method still has the following problems when it is applied to the detection of heterologous genomic DNA or heterologous cell-free DNA: the experimental operation of the NGS technical scheme is cumbersome, the detection period is long (3-7 working days), and the detection cost is high, so it is not suitable for Regular monitoring after transplantation; while other conventional technologies have disadvantages such as low throughput, many operation steps, low detection sensitivity, and easy pollution when opening the cover when detecting genetic polymorphism information

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  • Method and kit for detecting presence or proportion of donors in acceptor sample
  • Method and kit for detecting presence or proportion of donors in acceptor sample
  • Method and kit for detecting presence or proportion of donors in acceptor sample

Examples

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Embodiment 1

[0419] Embodiment 1. Selection of candidate SNP sites

[0420] The SNP site covered by the present invention is selected from the single nucleotide polymorphism site library (dbSNP) of the National Center for Biotechnology Information (NCBI), and the SNP site of the present invention preferably meets the following conditions: (1) Fst (population fixation coefficient) between different races 1Mb; (5) In order to avoid linkage between different loci, try to choose to be located on different chromosomes on the site. Carry out the screening of SNP locus according to above-mentioned standard, present embodiment has selected preferred 23 SNP loci, specifically as shown in table 1, SNP locus information and sequence are queried from the dbSNP database of U.S. National Center for Biotechnology Information (NCBI) And download, the allele frequency refers to the Asian population frequency from the Thousand Genomes database, and these sites are evenly distributed on each chromosome of...

Embodiment 2

[0424] Example 2. Determination of Chimeric Rate of Bone Marrow Transplant Donors

[0425] The detection process of this example is as follows figure 2 As shown, taking two bone marrow transplant sample groups as an example, the following two parts of samples were collected: 1. Collect and extract the donor samples and recipient samples of bone marrow transplant patients before transplantation for SNP typing, SNP typing principle such as figure 1 shown. 2. Collect peripheral blood at various time points during the recipient monitoring period after transplantation, and extract genomic DNA for quantification of target SNP sites, detection of donor chimerism after bone marrow transplantation, and evaluation of chimerism after allogeneic hematopoietic stem cell transplantation state.

[0426] The specific operation steps of the above detection process are as follows:

[0427] 1. Collect 2 bone marrow transplantation sample groups (each group includes donor samples before tr...

Embodiment 3

[0459] Example 3. The determination of the donor's free DNA ratio in organ transplantation (with donor information)

[0460] In this example, the determination of the proportion of donor cfDNA in the plasma and urine samples after a kidney transplantation was taken as an example to monitor the organ damage of kidney transplantation case 3, and investigate the feasibility of the method of the present invention for measuring the proportion of dd-cfDNA in organ transplantation and detection performance.

[0461] The detection process of this example case is as follows image 3 As shown, taking a kidney transplant sample group as an example, the following two parts of samples need to be collected: (1) Collect and extract the donor samples and recipient samples of kidney transplant patients before transplantation for SNP typing. Type principle such as figure 1 shown; or collect post-transplant recipient blood cell sediment, saliva, tissues other than transplanted organs, skin, ...

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Abstract

The invention relates to a method for detecting SNP sites of a donor-sourced sample and a receptor-sourced sample. Further, the present application also relates to a method for detecting the presence or proportion of donors in an acceptor sample, and a kit for implementing the method.

Description

technical field [0001] This application relates to the field of molecular diagnostics. Specifically, the present application relates to a method for detecting SNP sites in a donor-derived sample and a recipient-derived sample. Furthermore, the present application also relates to a method for detecting the presence or proportion of the donor in a recipient sample, and a kit for implementing the method. Background technique [0002] Heterologous DNA refers to the existence of non-self DNA from one or more individuals in the individual's own body. Compared with the individual's own DNA, the non-self DNA from one or more individuals can be defined as heterologous DNA. The most common example is the presence of donor-derived DNA in the recipient during allogeneic transplantation. The current detection method for heterologous DNA can be applied to two major aspects of bone marrow transplantation and solid organ transplantation. [0003] In bone marrow transplantation, allogeneic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6858C12Q1/6851C12Q1/6888C12N15/11
CPCC12Q1/6858C12Q1/6851C12Q1/6888C12Q2600/16C12Q2600/156C12Q2531/113C12Q2537/143C12Q2527/107C12Q2563/107C12Q2563/159C12Q2537/16C12Q2545/114C12N15/11C12Q1/6844
Inventor 李庆阁黄秋英陈昕雯
Owner XIAMEN UNIV
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