Group of HLA-related SNP marks and detection primer and determination method thereof

A certain method and marker technology, applied in the medical field, to achieve the effect of assisting the assessment of transplant rejection risk and autoimmune disease

Active Publication Date: 2018-05-04
广州博富瑞医学检验有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, there is no clinical report on the application of DQ promoter SNP analysis to organ transplantation. If the DQ promoter SNP can be analyzed to clarify the expression level of DQ protein in patients under immune status, it can be predicted by the pa

Method used

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  • Group of HLA-related SNP marks and detection primer and determination method thereof
  • Group of HLA-related SNP marks and detection primer and determination method thereof
  • Group of HLA-related SNP marks and detection primer and determination method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1 Obtaining a group of HLA-associated SNP markers

[0039] 1.1 Obtaining of blood samples in Example 1

[0040] Example 1 The blood samples were 6 blood samples randomly drawn from the pre-transplant whole blood samples of kidney transplant patients collected by the Organ Transplantation Department of the First Affiliated Hospital of Sun Yat-sen University for genomic DNA extraction.

[0041] 1.2 Example 1 Extraction of Genomic DNA from Blood Sample

[0042]In this experiment, TIANamp Blood DNAKit (catalogue number: DP348) was used to extract genomic DNA from the blood sample of Example 1. The specific steps are as follows:

[0043] (1) Gently invert and mix the whole blood samples of the six patients in Example 1, and draw 200ul from it into a new 1.5ml EP tube (if the DNA extraction concentration is too low, you can also centrifuge the blood sample at 500g for 5min to separate the layers, and take The buffy coat cells in the middle are 200ul for subsequen...

Embodiment 2

[0077] The difference between this embodiment and the embodiment is that the primers used in the amplification have the nucleotide sequences shown in SEQ ID NO: 1 and SEQ ID NO: 5; forward sequencing can be used during sequencing, and reverse sequencing can also be used. For sequencing, the forward sequencing primer is SEQ ID NO:1, and the reverse sequencing primer is SEQ ID NO:5.

Embodiment 3

[0079] The difference between this embodiment and the embodiment is that the primers used in the amplification are the nucleotide sequences shown in SEQ ID NO:3 and SEQ ID NO:5; forward sequencing can be used during sequencing, and reverse sequencing can also be used , during sequencing, the forward sequencing primer is SEQ ID NO:3, and the reverse sequencing primer is SEQ ID NO:5.

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Abstract

The invention provides a group of HLA-related SNP marks and a detection primer and a determination method thereof, and aims at providing a primer and a method for transplanting risk evaluation. The technical scheme is that the SNP marks comprise a first SNP mark, a second SNP mark and a third SNP mark, wherein the first SNP mark is a basic group A or T in the 32623514 position of the human No.6 chromosome; the second SNP marker is a basic group G or T in the 32623555 position of the human No.6 chromosome; the third SNP mark is a basic group A or T in the 32623560 position of the human No.6 chromosome; a primer pair for detecting the group of SNP markers in the claim 1 have the nucleotide sequences shown as SEQ ID NO:1 to 2 or SEQ ID NO:1 and 5 or SEQ ID NO:3 and 5 or SEQ ID NO:4 and 5. Thedetermination method of the HLA-related SNP sites comprises the following steps of (1) extracting the genome DNA of a host cell; (2) performing PCR amplification on the template DNA; purifying the PCR product SAP/Exon I; (3) respectively performing forward and/or reverse sequencing amplification on the purified PCR product by a sequencing primer; purifying the sequencing product; performing ABI 37030xl capillary electrophoresis sequencing so as to determine the SNP site and genotype.

Description

technical field [0001] The invention relates to the field of medical technology, in particular to an analysis of SNP markers related to HLADQ promoter, and the risk assessment of rejection after organ transplantation is predicted by this group of SNP markers. Background technique [0002] In the field of organ transplantation, various rejection reactions caused by humoral immunity mediated by HLA antibodies are the most critical factors affecting the survival of transplanted people / grafts. [0003] SNP (Single nucleotide polymorphism, single nucleotide polymorphism) mainly refers to the DNA sequence polymorphism caused by a single nucleotide variation at the genome level. [0004] In the human population, HLA (Human leukocyte antigen, human leukocyte antigen) has abundant polymorphisms. In organ transplantation, the HLA antigen in the endothelial cells of the donor organ is often used as the recognition antigen, causing a series of rejection reactions and affecting the organ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6881C12Q1/6869C12N15/11
CPCC12Q1/6869C12Q1/6881C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 姚远余小林王琛
Owner 广州博富瑞医学检验有限公司
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