Detection method for evaluating transplantation injury and rejection risk after liver transplantation

A detection method and liver transplantation technology, applied in the field of biomedicine, can solve the problems of influence of result accuracy, low accuracy and sensitivity, low sensitivity and specificity, etc., and achieve the effect of enhancing sensitivity and improving specificity and accuracy

Pending Publication Date: 2022-04-22
WUXI DIAGNOSTICS LAB SHANGHAI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, for the health monitoring of grafts after solid organ transplantation, blood is often used for liver function tests, or puncture needles are used to collect tissues for pathological examination, but these two methods have certain limitations. In terms of functional examination, the indicators obtained by routine blood drawing functional examination, such as creatinine, ALT, AST, bilirubin, etc., have low sensitivity and specificity, and cannot accurately reflect the health status of the graft.
[0005] For the current gold standard tissue biopsy, although it can directly reflect the health status of the graft, it has the following major disadvantages: 1. Invasive detection, which causes great pain to the patient and damages the graft; 2. Abnormal detection At this time, the substantial damage of the graft has already occurred, and the damage is often already serious. At this time, it is often too late for clinicians to save the graft; Accuracy has a great impact, so the use of puncture needles to collect pathological tissues for biopsy, the accuracy and sensitivity of this method is not high

Method used

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  • Detection method for evaluating transplantation injury and rejection risk after liver transplantation
  • Detection method for evaluating transplantation injury and rejection risk after liver transplantation
  • Detection method for evaluating transplantation injury and rejection risk after liver transplantation

Examples

Experimental program
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Effect test

Embodiment 1

[0037] Example 1 Primer and Probe Sequence

[0038] The corresponding relationship between primers and probes used in the present invention and the targeted SNP sites is shown in the table below:

[0039]

[0040]

[0041]

[0042]

[0043] The above primers were synthesized at Invitrogen Trading (Shanghai) Co.

Embodiment 2

[0044] Example 2 Using the detection method provided by the present invention to assess the risk of graft injury and rejection after liver transplantation

[0045] 1. Sampling and extracting sample DNA

[0046] In a specific embodiment, with EDTANa 2 Anticoagulant tubes were used to collect the preoperative venous blood of clinical transplant patients (recipients) and part of the transplanted organs of the donors, and the nucleic acid extraction kit (DNeasy Blood and Tissue kit, QIAGEN Company) was used to extract human genomic DNA from the whole blood of the recipients. and human genomic DNA in donor tissues were quantified by NanoDrop 2000 and normalized to 30ng / μL.

[0047] 2. qPCR detection

[0048] ABI 7500 fluorescent quantitative PCR instrument was used to perform single-hole amplification on the DNA region where the 39 SNP sites were located.

[0049] The PCR reaction system is shown in the table below, wherein the qPCR reaction reagent is TAKARA-RR392A, and the nucle...

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Abstract

The invention discloses a detection method for evaluating transplantation injury and rejection risks after liver transplantation, which comprises the following steps: 1) respectively adding primers and probes by taking genome DNA (Deoxyribose Nucleic Acid) of a donor source and genome DNA of a receptor source as templates; 2) performing qPCR detection on the DNA region where the SNP site is located; (3) respectively screening out 1 to 2 receptor homozygous SNP sites and donor homozygous SNP sites; 4) extracting free DNA of the receptor sample as a template, selecting a primer and a probe corresponding to the selected SNP site, adding the primer and the probe into the free DNA, and then carrying out ddPCR detection; and 5) analyzing the ddPCR result, and calculating the percentage of the free DNA containing the SNP site in the donor in the free DNA of the receptor sample. According to the detection method for evaluating the injury and rejection risk of the graft after the liver transplantation, the injury condition and rejection risk of the liver graft can be sensitively, specifically and accurately evaluated, any wound detection is not needed, the operation is convenient, the time is saved, and the efficiency is improved.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a detection method for evaluating graft damage and rejection risk after liver transplantation. Background technique [0002] Organ transplantation is the process of surgically transferring an organ in whole or in part from one individual to another. The aim is to replace a damaged or non-functional organ with a good one from a donor. The party providing the organ is the organ transplant donor, and the party receiving the organ is the organ transplant recipient. [0003] Transplant rejection (transplant rejection) is a problem that organ transplant patients need to be vigilant for life. Transplant rejection refers to that after the recipient undergoes allogeneic tissue or organ transplantation, the foreign tissue or organ is used as a kind of "alien component". Recognition by the recipient's immune system, which mounts an immunological response for attack, destruction and cl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/686C12N15/11
CPCC12Q1/6883C12Q1/686C12Q2563/107C12Q2545/114C12Q2563/159
Inventor 方焯刘釜均何赏尹晓莹
Owner WUXI DIAGNOSTICS LAB SHANGHAI CO LTD
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