JP-7857351-B2 - Methods for comparing genetic identity, and methods for determining the identity of individuals from which multiple samples originate.

JP7857351B2JP 7857351 B2JP7857351 B2JP 7857351B2JP-7857351-B2

Inventors

何鈞軒

Assignees

何鈞軒

Dates

Publication Date: 20260512
Application Date: 20240709
Priority Date: 20240131

Claims (20)

A method for comparing the genetic identity between the target gene group of a target sample and the test gene group of a test sample , A step of providing the target sample containing the aforementioned target gene group, A step of providing a test sample including a first test sample, a second test sample, and a third test sample, A step of comparing the identity between the human leukocyte antigen gene of the first test sample and the human leukocyte antigen gene of the target gene group, and obtaining the comparison result of the human leukocyte antigen gene, The process involves comparing the identity between the short tandem repeat sequence of the second test sample and the short tandem repeat sequences of the target gene group, and obtaining the comparison results of the short tandem repeat sequences. The process involves comparing the identity between the killer cell immunoglobulin-like receptor gene of the third test sample and the killer cell immunoglobulin-like receptor gene of the target gene group, and obtaining the comparison results for the killer cell immunoglobulin-like receptor genes. The step of determining that the target gene group is the same as the test gene group of the test sample if the comparison results of the human leukocyte antigen gene, the comparison results of the short tandem repeat sequence, and the comparison results of the killer cell immunoglobulin-like receptor gene are all the same, Methods that include...
The method according to claim 1, wherein the first test sample includes blood.
The step of comparing the identity between the human leukocyte antigen gene of the first test sample and the human leukocyte antigen gene of the target gene group is: The process involves extracting and amplifying the human leukocyte antigen gene from the first test sample, A step of using a next-generation sequencing method to compare the identity of the human leukocyte antigen gene in the first test sample with the human leukocyte antigen gene in the target gene group, The method according to claim 1, including the method described in claim 1.
The method according to claim 1, wherein the step of comparing the identity between the human leukocyte antigen gene of the first test sample and the human leukocyte antigen gene of the target gene group includes a step of comparing the identity of HLA-A, HLA-B, HLA-C, HLA-DRB1, HLA-DQB1, or combinations thereof.
The step of comparing the identity between the human leukocyte antigen gene of the first test sample and the human leukocyte antigen gene of the target gene group is: A step of comparing the identity between the coding region in the human leukocyte antigen gene of the first test sample and the coding region in the human leukocyte antigen gene of the target gene group, A step of comparing the identity between the non-coding region in the human leukocyte antigen gene of the first test sample and the non-coding region in the human leukocyte antigen gene of the target gene group, The method according to claim 1, including the method described in claim 1.
The method according to claim 1, wherein the second test sample comprises human tissue cells.
The step of comparing the identity between the short tandem repeat sequence of the second test sample and the short tandem repeat sequence of the target gene group is: The steps include extracting the short tandem repeat sequence from the second test sample, A step of using a sequencing typing method to compare the identity of the short tandem repeat sequence of the second test sample with the short tandem repeat sequence in the target gene group, The method according to claim 1, including the method described in claim 1.
The method according to claim 1, wherein the step of comparing the identity between the short tandem repeat sequence of the second test sample and the short tandem repeat sequence of the target gene group includes a step of comparing the identity of genes at the loci D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, the amelogenin gene, D5S818, FGA, or combinations thereof.
The method according to claim 1, wherein the third test sample includes blood.
The step of comparing the identity between the killer cell immunoglobulin-like receptor gene of the third test sample and the killer cell immunoglobulin-like receptor gene of the target gene group is as follows: The process involves extracting the killer cell immunoglobulin-like receptor gene from the third test sample, A step of comparing the identity of the killer cell immunoglobulin-like receptor gene of the third test sample with the killer cell immunoglobulin-like receptor gene in the target gene group, using a polymerase chain reaction-sequence-specific oligonucleotide method, a polymerase chain reaction-sequence-specific primer method, a sequencing typing method, or a combination thereof, The method according to claim 1, including the method described in claim 1.
The method according to claim 1, wherein the step of comparing the identity between the killer cell immunoglobulin-like receptor gene of the third test sample and the killer cell immunoglobulin-like receptor gene of the target gene group includes a step of comparing the identity of genes 2DL1, 2DL2, 2DL3, 2DL4, 2DL5, 2DS1, 2DS2, 2DS3, 2DS4, 2DS5, 3DL1, 3DL2, 3DL3, 3DS1, 2DP1, 3DP1, or combinations thereof.
A method for determining the identity between an individual from which a test sample originates and an individual from which a target sample originates , The steps include providing the aforementioned test sample and the aforementioned target sample, A step of comparing the identity of human leukocyte antigen genes between the test sample and the target sample, and obtaining the comparison results of human leukocyte antigen genes, A step of comparing the identity of the short tandem repeat sequences between the test sample and the target sample, and obtaining the comparison results of the short tandem repeat sequences. A step of comparing the identity of killer cell immunoglobulin-like receptor genes between the test sample and the target sample, and obtaining the comparison results of the killer cell immunoglobulin-like receptor genes, The step of determining that the individual from which the test sample originates and the individual from which the target sample originates are the same if the comparison results of the human leukocyte antigen gene, the comparison results of the short tandem repeat sequence, and the comparison results of the killer cell immunoglobulin-like receptor gene are all the same. Methods that include...
The method according to claim 12, wherein the test sample and the target sample include blood.
The step of comparing the identity of the human leukocyte antigen gene between the test sample and the target sample is as follows: A step of extracting and amplifying the human leukocyte antigen gene from the test sample and the human leukocyte antigen gene from the target sample, A step of using next-generation sequencing to compare the identity of the human leukocyte antigen gene of the test sample with the human leukocyte antigen gene of the target sample, The method according to claim 12, including the method described in claim 12.
The method according to claim 12, wherein the step of comparing the identity of the human leukocyte antigen gene between the test sample and the target sample includes a step of comparing the identity of the genes HLA-A, HLA-B, HLA-C, HLA-DRB1, HLA-DQB1, or combinations thereof.
The step of comparing the identity of the human leukocyte antigen gene between the test sample and the target sample is as follows: A step of comparing the identity between the coding region in the human leukocyte antigen gene of the test sample and the coding region in the human leukocyte antigen gene of the target sample, A step of comparing the identity of the non-coding region in the human leukocyte antigen gene of the test sample with the non-coding region in the human leukocyte antigen gene of the target sample, The method according to claim 12, including the method described in claim 12.
The step of comparing the identity of the short tandem repeat sequence between the test sample and the target sample is as follows: A step of extracting the short tandem repeat sequence from the test sample and the short tandem repeat sequence from the target sample, A step of comparing the identity of the short tandem repeat sequence of the test sample with the short tandem repeat sequence of the target sample using a sequencing typing method, The method according to claim 12, including the method described in claim 12.
The method according to claim 12, wherein the step of comparing the identity of the short tandem repeat sequences between the test sample and the target sample includes comparing the identity of genes at the loci D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, the amelogenin gene, D5S818, FGA, or combinations thereof.
The step of comparing the identity of the killer cell immunoglobulin-like receptor gene between the test sample and the target sample is as follows: A step of extracting the killer cell immunoglobulin-like receptor gene from the test sample and the killer cell immunoglobulin-like receptor gene from the target sample, A step of comparing the identity of the killer cell immunoglobulin-like receptor gene of the test sample with the killer cell immunoglobulin-like receptor gene of the target sample using a sequence-specific oligonucleotide probe method, a polymerase chain reaction-sequence-specific primer method, a sequencing- typeding method, or a combination thereof, The method according to claim 12, including the method described in claim 12.
The method according to claim 12, wherein the step of comparing the identity of the killer cell immunoglobulin-like receptor gene between the test sample and the target sample includes the step of comparing the identity of the genes 2DL1, 2DL2, 2DL3, 2DL4, 2DL5, 2DS1, 2DS2, 2DS3, 2DS4, 2DS5, 3DL1, 3DL2, 3DL3, 3DS1, 2DP1, 3DP1, or combinations thereof.

Description

This disclosure concerns methods for comparing genetic identity and methods for determining the identity of individuals from which multiple samples originate. In cell therapy or organ transplantation, the higher the genetic identity between the target cells and the original individual cells, the lower the risk of immune rejection. The risk of immune rejection can be minimized when the target cells and the original individual cells originate from the same individual. Therefore, if an individual is healthy, their cells can be cryopreserved in advance for use in future cell therapy or organ transplantation. However, because cell cryopreservations typically store large quantities of cells from different individuals simultaneously, misusing the target cells used for treatment can cause a serious immune rejection reaction in the individual. Therefore, to mitigate the risk of immune rejection, it is necessary to confirm whether the target cells (hereinafter referred to as the test sample) and the original individual cells (hereinafter referred to as the target sample) originate from the same individual before proceeding with treatment. However, conventional gene comparison methods have limited accuracy (for example, they identify short tandem repeat sequences (STRs), and if genes are 80% or more identical, they are judged to be the same individual), and there is still a significant risk of misidentification. To make the above and other objectives, features, advantages, and embodiments of the present invention clearer, the accompanying drawings are described below. This is a flowchart illustrating a method for comparing gene identity in several embodiments of the disclosed information. This is a flowchart illustrating a method for determining the identity of individuals from which multiple samples originate, in some embodiments of the present disclosure. To provide a more detailed and complete description of the present invention, embodiments and specific examples of the invention are described below in detail; however, these are not the only forms of carrying out or applying specific examples of the invention. Each of the embodiments disclosed below may be combined or substituted for one another without further description or explanation if beneficial, and other embodiments may be added to one embodiment. Numerous specific details are described in detail below to allow the reader to fully understand the following embodiments. However, embodiments of the invention can be carried out without these specific details. In this specification, unless otherwise specified in the text, the articles "one" and "the" generally refer to one or more. For further understanding, the terms "include," "contain," "have," and similar terms used herein specify the features, areas, integers, processes, operations, elements, and/or components described, but do not exclude other features, areas, integers, processes, operations, elements, components, and/or groups thereof. The following describes the methods disclosed herein using a series of operations or steps, but the procedures shown in these operations or steps should not be construed as limiting the invention. For example, some operations or steps may be performed in different procedures and/or simultaneously with other steps. Furthermore, it is not necessary to perform all operations, steps, and/or features to realize embodiments of the invention. Moreover, each operation or step described herein may include multiple sub-operations or steps. In this specification, polymerase chain reaction-sequence-specific oligonucleotide method (PCR-SSOP) refers to a method in which a probe labeled with an isotope or non-radioactive material is hybridized with fragments of a group of test genes amplified by polymerase chain reaction (PCR), and the genotype of the test gene group is determined via the signal resulting from the hybridization. In this specification, polymerase chain reaction-sequence-specific primer method (PCR-SSP) refers to a method that uses PCR to amplify genes with specific primers, and then determines the genotype of the tested gene group based on whether or not amplification has occurred. In this specification, sequencing-based typing (SBT) refers to a method of directly determining the sequence of a gene, such as Sanger SBT (SSBT) or next-generation sequencing (NGS). Several embodiments of this disclosure compare the identity of human leukocyte antigen (HLA) genes, short tandem repeat sequences (STR), and killer-cell immunoglobulin-like receptor (KIR) genes in test samples and target samples to confirm whether the target gene group is identical to the test gene group, and to determine whether the test sample and target sample originate from the same individual. By comparing highly polymorphic triple genes, it is as if the test sample and target sample are encrypted with a gene password lock, minimizing the probability of misidentifying the test sample (the probability of ident