Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Special primers for detecting SNP sites of drug-resistance related genes for anticoagulant and antiplatelet medication

An anti-platelet and drug-resistant technology, applied in the biological field, achieves good accuracy, is suitable for popularization and application, and reduces detection time

Active Publication Date: 2021-03-26
PEOPLES HOSPITAL PEKING UNIV
View PDF3 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no application of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) technology for precise drug genetic testing of anticoagulant and antiplatelet drug resistance

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Special primers for detecting SNP sites of drug-resistance related genes for anticoagulant and antiplatelet medication
  • Special primers for detecting SNP sites of drug-resistance related genes for anticoagulant and antiplatelet medication
  • Special primers for detecting SNP sites of drug-resistance related genes for anticoagulant and antiplatelet medication

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0101] This example is used to illustrate the special primers provided by the present invention for detecting SNP sites of genes related to drug resistance to anticoagulation and antiplatelet drugs.

[0102] According to the six commonly used anticoagulant and antiplatelet drugs: warfarin, aspirin, clopidogrel, dabigatran etexilate, rivaroxaban, and ticagrelor, the SNP sites related to precise drug use: rs730012, rs6065, rs4986893, rs4148738, rs1045642, rs113681054, rs4149056, rs4244285, rs28399504, rs1330344, rs1057910, rs2244613, rs2032582, rs9923231, rs8192935 were used to form a PCR reaction system with single-base specificity multiplex primers and nested primers were designed. Laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) technology (MASSARRAY platform) realizes the precise drug early warning detection of the above-mentioned 6 drugs used for anticoagulant and antiplatelet therapy.

[0103] Specific PCR primers and single base extension primer...

Embodiment 2

[0150] This example is used to illustrate the method provided by the present invention for detecting SNP sites of genes related to drug resistance to anticoagulation and antiplatelet drugs.

[0151] 1. Template whole genome DNA extraction

[0152] Genomic DNA was extracted from peripheral blood of 384 cases, and the concentration was uniformly diluted to 20-30ng / μL.

[0153] 2. PCR reaction

[0154] Prepare a reaction system in a 5 μL PCR 96-well plate: 1 μL template DNA, 1 μL amplification primer Mix, PCR buffer (containing 15 mM Mg 2+ )0.5μL, MgCl 2 (25mM) 0.4μL, dNTP (25mM) 0.1μL, Hotstar Taq (5U / μL) 0.2μL, make up to 5μL with MBG (Molecular Biology Grade) water. Seal the 384-well plate with Parafilm.

[0155] The PCR amplification primer Mix consists of primers shown in sequence 1-30 and water, wherein the concentration of each primer is 0.5 μM.

[0156] The above PCR reaction program is set as follows:

[0157]

[0158] 3. Alkaline phosphatase treatment (SAP dige...

Embodiment 3

[0185] This example is used to illustrate the method for detecting the SNP sites of 6 anticoagulant and antiplatelet drug resistance-related genes and the application of special primers.

[0186] 1. Template DNA (whole genome) extraction

[0187] The peripheral blood of 8 subjects was taken as samples 1-8 to be tested (corresponding to subjects 1-8 in turn), and subjects 1-8 were all diagnosed with hypertension, coronary heart disease (stable angina, unstable angina) or old myocardial infarction), and peripheral atherosclerotic plaque.

[0188] 2. PCR reaction: see step 2 of Example 2.

[0189] 3. Alkaline phosphatase treatment (SAP digestion reaction): see step 3 of Example 2.

[0190] 4. Single base extension reaction: refer to step 4 of Example 2.

[0191] 5. Resin purification: refer to Step 5 of Example 2.

[0192] 6. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry detection: see step 6 of Example 2.

[0193] After testing, in the 8 teste...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to the technical field of biology, and provides a special primer for detecting SNP sites of drug-resistance related genes for anticoagulant and antiplatelet medication. The special primer comprises a primer group 1 and a primer group 2, wherein the primer group 1 is a specific PCR primer composed of 15 pairs of primers, and the primer group 2 is a single base extension primercomposed of 15 primers. The invention further provides a reparation for detecting the SNP sites of drug-resistance related genes for anticoagulant and antiplatelet medication, and a method for detecting the SNP sites of the drug-resistance related genes for anticoagulant and antiplatelet medication. By adopting the special primer provided by the invention, the detection time can be greatly shortened, the cost is saved, and the special primer has a very high application value for guiding precise medication of six clinically common anticoagulant and antiplatelet drugs, and is suitable for popularization and application.

Description

technical field [0001] The invention belongs to the field of biotechnology, and particularly relates to a special primer for detecting the SNP site of a gene related to anticoagulant and antiplatelet drug resistance and its application. Background technique [0002] Aspirin, clopidogrel, and ticagrelor are commonly used antiplatelet drugs in clinical practice. Aspirin is the basic drug for the prevention and treatment of arterial thrombosis; clopidogrel is a platelet aggregation inhibitor, which is used to prevent and treat heart, brain and other arterial circulation disorders caused by high platelet aggregation, such as preventing the formation of thrombus; Grerelor is a new type of anti-platelet drug similar to clopidogrel, which can directly inhibit the P2Y12 receptor without metabolic activation, and can inhibit platelet aggregation more effectively than clopidogrel; at the same time Its inhibition on platelets is reversible, and the degree of inhibition reflects the pl...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/156C12Q2600/106Y02A50/30
Inventor 赵美封宇飞黄琳郑方芳宋俊贤
Owner PEOPLES HOSPITAL PEKING UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com