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Gene detection method, primer probe combination and kit for precise drug use of depression

A precision medicine and gene detection technology, applied in the direction of recombinant DNA technology, DNA / RNA fragments, etc., can solve the problem of inaccurate prediction of drug efficacy and adverse reactions, etc.

Inactive Publication Date: 2019-06-25
广州海思医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, there is no attention or detection of related genes other than drug metabolizing enzymes, and the prediction of drug efficacy and adverse reactions is not accurate enough

Method used

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  • Gene detection method, primer probe combination and kit for precise drug use of depression
  • Gene detection method, primer probe combination and kit for precise drug use of depression
  • Gene detection method, primer probe combination and kit for precise drug use of depression

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The design of embodiment 1 primer and probe

[0034] According to the metabolism and efficacy of commonly used antidepressants in the body, the genes and loci that affect the efficacy and adverse reactions of the drugs are determined. Related genes include CYP2D6, CYP2C19, FKBP5, HTR2A, and HTR1A.

[0035] Based on the genes listed above, 11 different gene loci are involved, and the specific loci are as follows:

[0036] serial number

Gene

type

rs number

1

CYP2D6

c.2850C>T(*2, *41)

rs16947

2

CYP2D6

c.1846G>A(*4)

rs3892097

3

CYP2D6

c.100C>T(*4, *10)

rs1065852

4

CYP2D6

c.4180G>C(*2, *4, *10, *41)

rs1135840

5

CYP2D6

c.2988G>A(*41)

rs28371725

6

CYP2C19

c.681G>A(*2)

rs4244285

7

CYP2C19

c.636G>A(*3)

rs4986893

8

CYP2C19

c.-806C>T(*17)

rs12248560

9

FKBP5

c.-20+18122T>C

rs4713916

10

HTR2A

c.614-...

Embodiment 2

[0042] The preparation of embodiment 2 kit

[0043] A kit for simultaneous detection of CYP2D6 / CYP2C19 / FKBP5 / HTR2A / HTR1A gene polymorphisms, comprising 11 tubes of PCR reaction solution for respectively detecting gene locus polymorphisms and 2 tubes of quality control products. According to the experimental exploration, the usage of each component of the PCR reaction system is as follows:

[0044] components

Dosage (μL)

PCR reaction buffer

5

Upstream primer (F) (10 μM)

0.1-1.0

Downstream primer (R) (10μM)

0.1-1.0

Wild type detection probe (10μM)

0.1-1.0

Mutant detection probe (10μM)

0.1-1.0

dUTP (10mM)

0.1

UNG enzyme (0.1U / μL)

1

Sample DNA (>0.5ng / μL)

1-5

Sterilized purified water

make up to 20

[0045] PCR reaction buffer, using Thermo Fisher Scientific (China) Co., Ltd. Genotyping Master Mix reagents.

[0046]The volume of each primer and probe is: 0.4 μL for rs169...

Embodiment 3

[0048] The use of embodiment 3 kits

[0049] The present invention can be used to detect various samples such as blood, buccal swab, dried blood film, etc. Now, two cases of anticoagulated blood samples are taken as examples to illustrate the specific use of the kit in Example 2.

[0050] (1) Sample DNA extraction

[0051] Take 1-2ml of anticoagulant blood samples from 2 cases, use the blood genomic DNA extraction kit (spin column type) produced by Tiangen Biochemical Technology (Beijing) Co., Ltd. (article number: DP318 specification: 50 / 200 servings), follow the Operation for DNA extraction.

[0052] The DNA concentration (≥0.5 ng / μL) and purity (1.6-2.0) of the extracted DNA were measured using a micro-volume ultraviolet spectrophotometer. The sample DNA concentration was adjusted to 5-25ng / μL, which was used as the template to be tested.

[0053] (2) Preparation of reaction system

[0054] Take 18 μL of the detection reaction solution into PCR tubes, add 2 μL of sample...

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Abstract

The invention relates to the field of gene detection, in particular to a gene detection method, a primer probe combination and a kit for precise drug use of depression to overcome the shortcoming thatthe market is short of products for gene detection of the drug use of the depression, only detection of a drug metabolism gene is adopted clinically, and the drug therapeutic effect predication is inaccurate. 11 loci of 5 related genes of CYP2D6, CYP2C19, FKBP5, HTR2A and HTR1A of anti-depression drugs are detected to determine the relevant gene locus types, and the clinical drug use can be guided by analysis.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a gene detection method for precise medication for depression, a combination of primers and probes, and a kit. Background technique [0002] Depression, also known as depressive disorder, is a common mental illness. According to statistics from the World Health Organization, by 2020, depression will become the second leading disease after heart disease. At present, more than 300 million people in the world are suffering from depression. The prevalence rate of depression in China is 6.1%, which is 3 percentage points higher than the global average, and the incidence rate has been increasing year by year in recent years. It is converted according to the incidence rate of 6.1%. , domestic depression patients have reached 90 million. [0003] Currently, the most effective means of treating depression is drug therapy, but drug efficacy and adverse reactions show significant individual ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
Inventor 夏昊强周煌凯王芳刘倩关丽雅邓美英陈秋红陆嫚云曹治家
Owner 广州海思医疗科技有限公司
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