A kit for simultaneously detecting statin metabolizing gene multisite mutations

A multi-site mutation and kit technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of low lipid-lowering effect and detection kits that are not released by any manufacturer, and achieve the effect of high sensitivity

Inactive Publication Date: 2017-08-29
NINGBO MEIJING MEDICAL TECH
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AI Technical Summary

Problems solved by technology

[0007] (4) ABCB1 multi-drug resistance gene 1, the expressed protein is a drug transporter, and there are two SNP polymorphisms in the gene sequence that have certain effects on the metabolism of statins, which are genome 2677 and 3435 respectively, Both of these loci are mutant individuals with less lipid-lowering effect when using atorvastatin
[0009] At present, there are related kits for the detection of polymorphisms of the statin drug metabolism genes APOE and SLCO1B1 in China, but no detection kits for the five genes (APOE, SLCO1B1, CETP, ABCBI, MTHFR) have yet been released by any manufacturer

Method used

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  • A kit for simultaneously detecting statin metabolizing gene multisite mutations
  • A kit for simultaneously detecting statin metabolizing gene multisite mutations
  • A kit for simultaneously detecting statin metabolizing gene multisite mutations

Examples

Experimental program
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Effect test

Embodiment 1

[0047] 1. Nucleic acid DNA extraction

[0048] 1.1 Add 200ul of whole blood sample to be tested, 20ul of proteinase K, and 400ul of lysate to a 1.5ml centrifuge tube in turn, vortex and mix well, and place in a 56°C water bath or metal bath for 10 minutes.

[0049] 1.2 Add 400ul of isopropanol, vortex for 10 seconds to mix well, then add 20ul of magnetic beads, vortex for 10 seconds, and let stand at room temperature for 10 minutes.

[0050] 1.3 Put the centrifuge tube on the magnetic separation rack and let it stand for 10 seconds until all the magnetic beads are adsorbed to the wall of the centrifuge tube (if there is liquid on the centrifuge tube cover, keep the centrifuge tube upside down on the magnetic separation rack for 3-5 times, Make the magnetic beads on the cap of the tube also be adsorbed to the tube wall), carefully suck off all the liquid, and be careful not to touch the magnetic beads.

[0051] 1.4 Remove the centrifuge tube, add 750ul washing solution I, vortex...

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Abstract

The invention relates to a kit for simultaneously detecting statin metabolizing gene multisite mutations. The kit includes primer pairs and probe pairs for detecting APOE, SLCO1B1, CETP, ABCB1 and MTHFR gene sites. MGB (minor groove binder) probes and a real-time fluorescent PCR technique are applied in the kit. The kit has advantages of capability of being time saving and convenient, high sensitivity, capability of allowing positive and negative coincidence rates of a sample to be 99.5% or above, and the like. The kit is mainly used for personalized medication assisted diagnosis of statins such as simvastatin, atorvastatin and pravastatin.

Description

technical field [0001] The invention belongs to the field of statin drug detection, in particular to a kit for simultaneously detecting multi-site mutations of statin drug metabolism genes. Background technique [0002] Statins are widely used lipid-lowering drugs, which can effectively prevent and treat cardiovascular and cerebrovascular diseases. In the United States, more than 25% of people older than 45 years old take statins. Especially in the secondary prevention of coronary heart disease after percutaneous stent implantation (PCI), the use of active statin lipid-lowering therapy is of great significance for the prevention and treatment of coronary heart disease. [0003] As the dose of statins increases, the effect of lowering LDL is enhanced, but at the same time, the incidence of adverse reactions is also high. One of the main toxic and side effects of statins is myopathy, and 5-29% of patients will develop statin-induced myopathy (SIM), which mainly manifests as f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q1/6883C12Q2600/106C12Q2600/156C12Q2600/16C12Q2561/101C12Q2545/113C12Q2561/113
Inventor 张晓晶沈挺李春燕宋毅
Owner NINGBO MEIJING MEDICAL TECH
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