Application and detection kit of SNP sites

A kit and site technology, applied in the biological field, can solve problems such as low accuracy and sensitivity, and the impact on the accuracy of results

Active Publication Date: 2017-08-18
成都仕康美生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] 3) If the puncture needle does not collect the lesion tissue part of the graft, it will have a great impact on the accuracy of the results, so the accuracy and sensitivity are not high

Method used

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  • Application and detection kit of SNP sites
  • Application and detection kit of SNP sites
  • Application and detection kit of SNP sites

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Using the kit provided by the present invention to assess the risk of graft injury and rejection after liver transplantation

[0044] Extraction of total free DNA from plasma

[0045] 1. Use Streck non-invasive blood collection tube to collect 10ml of blood from a recipient 2 weeks after liver transplantation;

[0046] 2. Transfer the blood collected from the Streck non-invasive blood collection tube to a 15ml centrifuge tube, and centrifuge at 2000g for 10 minutes at a low temperature of 4℃; take about 4-6ml of the upper plasma in a new 15ml centrifuge tube, continue to centrifuge at 4℃ for 10 minutes at 4000g; take the upper plasma 4~6ml in a new 15ml centrifuge tube;

[0047] 3. Take 4ml of plasma and use QIAamp Circulating Nucleic Acid Kit from QIAGEN to extract total free DNA, adjust the DNA concentration to 10-40ng / ul.

[0048] Preparation of digital PCR reaction droplets

[0049] 1. Configure a digital PCR reaction system, prepare digital PCR reaction droplets, a...

Embodiment 2

[0067] An EDTA blood collection tube was used to collect 2 ml of blood from the recipient 2 weeks after liver transplantation in Example 1, and routine liver function tests were performed. The results are shown in Table 2.

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Abstract

The invention relates to the technical field of biology, in particular to application and a detection kit of SNP sites. The invention provides the application of the 60 SNP sites to assessment on post-liver transplantation graft injury and rejection risk; through detection on the specific 60 SNP sites of total receptor free DNA after transplantation and combination with a specific calculating method, the content of donor free DNA in the receptor free DNA, namely GcfDNA donor and receptor ratio is finally obtained; the degree of graft rejection is quickly and accurately judged according to the GcfDNA donor and receptor ratio without any intrusive detection, and a simple and effective auxiliary means is provided for clinical detection on the post-organ transplantation graft injury and the rejection risk.

Description

Technical field [0001] The invention relates to the field of biotechnology, in particular to the application of SNP sites and a detection kit thereof. Background technique [0002] At present, the health monitoring of transplants after solid organ transplantation usually adopts blood drawing for liver function examination, or puncture needle to collect tissue for pathological examination. [0003] For routine blood sampling function tests, the sensitivity and specificity of various indicators such as creatinine, ALT, AST, bilirubin, etc. are not high, and they cannot accurately reflect the health of the graft. [0004] For the current gold standard tissue biopsy, it can directly reflect the health of the graft. But there are the following major disadvantages: [0005] 1) Invasive detection, causing great pain to the patient and damage to the graft at the same time; [0006] 2) When the abnormality is detected, substantial damage to the graft has occurred, and the damage is often sever...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q2600/156
Inventor 魏亮卢俊
Owner 成都仕康美生物科技有限公司
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