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Primer group for detecting hypertension medicine metabolism related genes and reagent kit

A primer set and high blood pressure technology, applied in the fields of medicine and biology, can solve the problems of long detection cycle, cumbersome operation, uncontrollable factors of test results, etc., achieve high sensitivity and specificity, simple sample processing, and reduce adverse drug reactions Effect

Active Publication Date: 2018-12-25
PRECEDO PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are many methods for detecting gene mutations, the most commonly used are polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP method), sequence-specific PCR, manual or automatic sequencing, gene chip (gene chip, Also known as DNA chips), these methods are cumbersome to operate, the detection cycle is long, and the factors affecting the test results are uncontrollable, so they are currently only used for scientific research

Method used

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  • Primer group for detecting hypertension medicine metabolism related genes and reagent kit
  • Primer group for detecting hypertension medicine metabolism related genes and reagent kit
  • Primer group for detecting hypertension medicine metabolism related genes and reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] Embodiment 1: the preparation of kit

[0098] 1. Design and synthesis of primers for genes related to hypertension drug metabolism

[0099] Using oligo software to design BDKRB2 gene, CYP11B2 gene, OAT1 / OAT3 gene, AGT gene, KCNH2 gene, LDLR gene, APOB gene, CACNA1C gene, GRK4 gene, ACE gene, CYP2D6 gene, CYP2C9 gene, CYP2C19 gene, ACE2 gene, AGTR1 Gene, ADRB2 gene and NR3C2 gene, the PCR amplification primers and sequencing primers of 17 hypertensive drug metabolism-related genes (see Table 1). The primer design follows the following principles: the length of the primer is about 25-30bp; the primer has no dimer; no hairpin structure; the Tm value difference between the upstream and downstream primers should not exceed 3°C; the GC content is about 50%. After the upstream and downstream primers are determined, use Blast to compare and analyze the upstream and downstream primers in the NCBI database to ensure the specificity and amplification efficiency of the primers; ...

Embodiment 2

[0106] Embodiment 2: the use of kit

[0107] 1. Sample testing

[0108] Take out the relevant reagents, and prepare the corresponding system according to the amount of template as follows: Take the PCR reaction solution, mix it evenly, add sample DNA, negative control substance or positive control substance as template to form the reaction system. Perform PCR amplification according to the PCR reaction procedure.

[0109] The main components of each gene detection system for hypertension are shown in Table 3.

[0110] Table 3. Main components of each gene detection system for hypertension

[0111]

[0112] The PCR reaction program of this system is shown in Table 4.

[0113] Table 4. PCR reaction program

[0114]

[0115] After the amplification is completed, agarose gel electrophoresis is performed to check the PCR amplification product, and the PCR amplification product is purified before proceeding to the next step.

[0116] 2. Sanger sequencing

[0117] Follo...

Embodiment 3

[0133] Example 3: The effect of the primer set and / or kit of the present invention on the medication guidance of hypertensive patients

[0134] In order to verify the effect of the primer set and / or kit of the present invention on the drug guidance effect of hypertensive patients, volunteers with hypertension were selected at the "Present Medical Laboratory", and the genotype was performed using the invented primer set and / or kit. Test, according to the test results of each volunteer, accurately formulate its medication plan, and conduct clinical follow-up to observe the clinical drug effect.

[0135] Medication Guidance Example 1

[0136] The tester Tian, ​​who took spironolactone before the test, did not have a good effect. Using the primer set and / or kit of the present invention, Tian was tested for genes related to hypertension. According to Tian's gene detection results, he predicted hydrochlorothiazide and Propranolol is more effective. After Tian took propranolol ...

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Abstract

The invention relates to a primer group for detecting hypertension medicine metabolism related genes on the basis of Sanger sequencing methods and a reagent kit with the primer group. The primer groupis characterized by comprising primers for detecting at least one type of genes selected from BDKRB2 genes, CYP11B2 genes, OAT1 / OAT3 genes, AGT genes, KCNH2 genes, LDLR genes, APOB genes, CACNA1C genes, GRK4 genes, ACE genes, CYP2D6 genes, CYP2C9 genes, CYP2C19 genes, ACE2 genes, AGTR1 genes, ADRB2 genes and NR3C2 genes.

Description

technical field [0001] The invention relates to the fields of medicine and biotechnology, in particular to a primer set for detecting genes related to hypertension drug metabolism based on Sanger sequencing method, a kit using the primer set, and the use of the primer set in preparing the kit. Background technique [0002] Hypertension is a worldwide problem, and it cannot be cured at present. According to statistics, about 1 / 3 of the adults in the world suffer from high blood pressure. Hypertensive patients are prone to heart disease, stroke, chronic kidney disease and premature death, which pose a serious threat to human health and life. At present, the number of hypertensive patients in my country has reached 330 million, and the incidence rate is increasing year by year; drugs are the main means of treating hypertension and its complications at present and for a long time in the future. Currently, the five major classes of antihypertensive drugs are calcium channel bloc...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/106C12Q2600/156
Inventor 刘娟周玲燕张定芳
Owner PRECEDO PHARMA CO LTD
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