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Application of snp site and its primers, probes and detection kits

A site and reagent technology, which can be used in the determination/inspection of microorganisms, recombinant DNA technology, biochemical equipment and methods, etc., and can solve problems such as low accuracy and sensitivity, and impact on the accuracy of results

Active Publication Date: 2021-01-12
成都仕康美生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 primers, probes and detection kits
  • Application of snp site and its primers, probes and detection kits
  • Application of snp site and its primers, probes and detection kits

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0126] Example 1 Using the kit provided by the present invention to assess the risk of graft injury and rejection after renal transplantation Extraction of total free DNA in plasma

[0127] 1. Using Streck non-invasive blood collection tubes to collect 10ml of blood from a recipient 2 weeks after kidney transplantation;

[0128] 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;

[0129] 3. Take 4ml of plasma, use QIAamp Circulating Nucleic Acid Kit from QIAGEN to extract the total free DNA, and adjust the DNA concentration to 10-40ng / ul.

[0130] Preparation of Digital PCR Reaction Droplets

[0131] 1. Configure a digital PCR reaction system, prepare digital PCR...

Embodiment 2

[0162] 2 ml of blood from the recipient 2 weeks after the kidney transplantation in Example 1 was collected using an EDTA blood collection tube, and routine renal function testing was performed. The results are shown in Table 5.

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Abstract

The invention relates to the technical field of biology, in particular to applications of SNP (single nucleotide polymorphism) sites as well as primers, probes and a detection kit for the SNP sites, and particularly provides applications of 40 SNP sites in evaluation of transplant damage and rejection risk after renal transplantation. The 40 specific SNP sites of total free DNA of a receptor after transplantation are detected, and by means of combination with a specific calculation method, content of free DNA of a donor in free DNA of the receptor is obtained finally, that is, GcfDNA donor-receptor ratio is obtained; the degree of transplant rejection is judged rapidly and accurately according to the GcfDNA donor-receptor ratio, any intrusive detection is not needed, and one simple and effective auxiliary means is provided for clinical detection of the transplant damage and rejection risk after organ transplantation.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of SNP sites and primers, probes and detection kits thereof. Background technique [0002] At present, the health monitoring of grafts after solid organ transplantation often uses blood drawing for renal function tests, or puncture needles for tissue collection 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 gra...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q1/6883C12Q2600/156C12Q2531/113C12Q2563/159
Inventor 魏亮
Owner 成都仕康美生物科技有限公司
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