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Application of snp site and its detection kit

A kit and site technology, applied in the biological field, can solve the problems of the influence of the accuracy of the results, low accuracy and sensitivity, etc.

Active Publication Date: 2018-04-10
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 of snp site and its detection kit
  • Application of snp site and its detection kit
  • Application of snp site and its detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Using the kit provided by the present invention to assess graft damage and rejection risk after liver transplantation Extraction of total free DNA in plasma

[0043] 1. Using Streck non-invasive blood collection tubes to collect 10ml of blood from a recipient 2 weeks after liver transplantation;

[0044] 2. Transfer the blood collected by the Streck non-invasive blood collection tube to a 15ml centrifuge tube, and centrifuge at 2000g at 4°C for 10min at low temperature; take about 4-6ml of the upper plasma in a new 15ml centrifuge tube, and continue to centrifuge at 4000g at 4°C for 10min at low temperature; take the upper plasma 4-6ml in a new 15ml centrifuge tube;

[0045] 3. Take 4ml of plasma, use the QIAamp Circulating Nucleic Acid Kit from QIAGEN to extract the total free DNA, and adjust the DNA concentration to 10-40ng / ul.

[0046] Preparation of Digital PCR Reaction Droplets

[0047] 1. Configure the digital PCR reaction system, prepare digital PCR rea...

Embodiment 2

[0066] 2 ml of blood from the recipient 2 weeks after liver transplantation in Example 1 was collected using EDTA blood collection tubes, 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 detection kits thereof. Background technique [0002] At present, the health monitoring of grafts after solid organ transplantation often uses blood drawing for liver function tests, or tissue collection by puncture needles for pathological examination. [0003] For routine blood drawing function tests, the sensitivity and specificity of various indicators such as creatinine, ALT, AST, bilirubin, etc. are not high, and cannot accurately reflect the health status of the graft. [0004] For the current gold standard tissue biopsy, although it can directly reflect the health status of the graft. But there are the following major disadvantages: [0005] 1) Invasive detection, which causes great pain to the patient and damages the graft at the same time; [0006] 2) When the abnormality is detected, the substantial damage of the graft has already occurr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883
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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