Gene polymorphism detection primer set, kit and detection method of caffeine metabolic capacity related gene CYP1A2

A CYP1A2, gene polymorphism technology, used in biochemical equipment and methods, recombinant DNA technology, and microbial assay/inspection, etc. problem, to achieve the effect of short time, not easy to contaminate, and high sensitivity

Inactive Publication Date: 2019-07-19
大连美纳医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

To put it simply, if you carry the A allele (wild) at the rs762551 site, you will easily have a stronger ability to metabolize caffeine. Caffeine can be metabolized in a short time, so the effect of caffeine is very strong. Limited, cannot produce a particularly obvious sense of excitement
Conversely, for people who carry the C allele (mutation), the clearance rate of caffeine in the body is very slow, and it takes a long time to take effect. Such people often have a cup of coffee that will make them sleepless at night, and some will also affect their sleep quality. Appetite, vomiting, and cramps, and adverse reactions such as gastritis or heart disease may also occur
At present, there is no comprehensive and specific detection method that can effectively detect this gene.

Method used

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  • Gene polymorphism detection primer set, kit and detection method of caffeine metabolic capacity related gene CYP1A2
  • Gene polymorphism detection primer set, kit and detection method of caffeine metabolic capacity related gene CYP1A2
  • Gene polymorphism detection primer set, kit and detection method of caffeine metabolic capacity related gene CYP1A2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A primer set for detecting polymorphism of CYP1A2 gene related to caffeine metabolism capacity, including the following primers:

[0029] Detection primer 1: 5'-AAGGGTGAGCTCTGTGGCTC-3' (SEQ ID NO.1);

[0030] Detection primer two: 5'-AAGGGTGAGCTCTGTGGCTA-3' (SEQ ID NO. 2);

[0031] Detection primer three: 5'-CCAGGCAGAAGATGGCAGAG-3' (SEQ ID NO.3);

[0032] Internal control primer 1: 5'-AGCAAGCAGGAGTATGACG-3' (SEQ ID NO.4);

[0033] Internal control primer 2: 5'-GAAAGGGTGTAACGCAACT-3' (SEQ ID NO. 5).

[0034] The detection primers and internal control primers described in this embodiment are all synthesized by biotechnology commonly used in the art.

Embodiment 2

[0036] Such as figure 1 As shown, a kit for detecting the polymorphism of the CYP1A2 gene related to caffeine metabolism capacity includes the five primers described in Example 1. The kit also includes a PCR reaction premix, a wild plasmid, a C5246A mutant plasmid and purified water which are respectively contained in a container. The PCR reaction premix used in this embodiment is a product sold by Tiangen Biochemical Technology (Beijing) Co., Ltd. The PCR reaction premix includes 3'→5' exonuclease activity high-fidelity Taq enzyme, 1.0-5.0mM MgCl2, 1.0-5.0mM dNTPs and SYBR Green I.

[0037] The kit includes a box body 1, the inside of the kit is divided into two independent cavities by a partition 3, and both cavities are provided with a sponge 2, the detection primer 1, the detection primer 2, the detection primer 3. Primer one, internal control primer two, PCR reaction premix, wild plasmid, C5246A mutant plasmid and purified water are respectively installed in the test tube, ...

Embodiment 3

[0040] A method for detecting the CYP1A2 gene polymorphism of the caffeine metabolism-related gene, using the detection primer 1 and the detection primer 3 of the present invention to detect the wild gene at the rs762551 site of the CYP1A2 gene, and using the detection primer 2 of the present invention And detection primers three pairs of CYP1A2 gene rs762551 mutation gene detection.

[0041] The method for detecting the polymorphism of the CYP1A2 gene related to caffeine metabolism capacity includes the following steps:

[0042] a. Mix the PCR reaction premix, detection primer one, detection primer three and water evenly to obtain reaction mixture one; mix the PCR reaction premix, detection primer two, detection primer three and water evenly to obtain reaction mixture two; Mix the PCR reaction premix, internal control primer one, internal control primer two and water evenly to obtain reaction mixture three;

[0043] b. Add the sample DNA to be tested into reaction mixture one, reac...

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Abstract

The present invention discloses a gene polymorphism detection primer set, a kit and a detection method of a caffeine metabolic capacity related gene CYP1A2. The gene detection primer set comprises three detection primers and two internal control primers. The present invention provides the primer composition for detecting CYP1A2 gene mutations of samples of fresh blood, etc., and when the primer composition is used for detecting a C5246A mutation of the CYP1A2 gene, the primer composition has advantages of high sensitivity, strong specificity, short time consumption, etc. The kit is simple instructure design and convenient to use. The partition design enables the detection process to be less susceptible to contamination and detection results are more accurate. The detection method is short in time consumption, high in sensitivity and strong in specificity.

Description

Technical field [0001] The invention belongs to the technical field of gene detection, and specifically relates to a primer set, a kit and a detection method for detecting the CYP1A2 gene polymorphism of a gene related to caffeine metabolism. Background technique [0002] The metabolism of caffeine in the body is mainly due to cytochrome P450 (CYP450, a superfamily of heme-thiolate proteins, which participates in endogenous substances and exogenous substances including drugs and environmental compounds). Substance metabolism) CYP1A2, a member of the family, is involved. There are a series of SNP sites in the gene that controls this enzyme, and their nucleotide mutations will cause each person's caffeine metabolism rate to be different. To put it simply, there is an A allele (wild) at the locus of rs762551, which will easily have a stronger ability to metabolize caffeine. Caffeine can be metabolized in a short time, so the effect of caffeine is very effective. It is limited and ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 田晓丽
Owner 大连美纳医学检验实验室有限公司
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