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Kit and method for detecting human pressure sensitivity genotypes

A genotype and kit technology, applied in the field of kits for detecting human stress sensitivity genotypes, can solve the problems of complicated operation of Sanger sequencing method, immature application of SNP detection technology and high cost

Inactive Publication Date: 2021-06-08
NINGBO HEALTH GENE TECHNOLOGIES CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the Sanger sequencing method is complex and expensive
When there are many sequencing sites, it needs to be sequenced one by one, which takes a long time and the cumulative price is relatively expensive
The next-generation sequencing technology realizes sequencing while synthesizing, which has the advantages of high throughput and high efficiency. However, the next-generation sequencing platform is expensive and has low popularity, and its clinical application as a SNP detection technology is not mature enough.
[0014] At present, there is no report on the multiple gene detection scheme related to pressure sensitivity based on multiple PCR and capillary electrophoresis separation technology in China

Method used

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  • Kit and method for detecting human pressure sensitivity genotypes
  • Kit and method for detecting human pressure sensitivity genotypes
  • Kit and method for detecting human pressure sensitivity genotypes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] (1) Selected sample: DNA / RNA-free water.

[0053] (2) System configuration

[0054] According to the instructions, prepare the primer Primer Mix and PCR reaction solution on ice according to the ratio of the instructions to configure the reaction system. After vortexing and mixing, centrifuge with a centrifuge, and then use a pipette tip to mix and distribute.

[0055] (3) Add samples

[0056] According to the instructions, use a pipette to take the corresponding volume of DNA / RNA-free water and add it to the reaction system that has been aliquoted.

[0057] (4) Amplification program

[0058] The amplification program on the PCR instrument is shown in Table 2.

[0059] (5) The amplified product is detected on the 3500DX genetic analyzer

[0060] The sample loading mixture is composed of deionized formamide and molecular weight internal standard (Size-500) in the system {(1 μL Size-500+12 μL deionized formamide)×(injection number)}. Mix 9 μL of the loading mixture w...

Embodiment 2

[0066] (1) Selected samples: exfoliated cell samples from the oral cavity wall of subject 1 and subject 2.

[0067] (2) Sample collection

[0068] Collection type: Exfoliated cells from the inner wall of the oral cavity.

[0069] Collection method: Use the saliva collection stick to wipe back and forth on the inner wall of the oral cavity 4 times, continue to wipe back and forth on the inner wall of the oral cavity 4 times with the reverse side of the saliva collection stick, take out the saliva collection stick, and press repeatedly on the saliva sample collection card to collect the saliva inner wall cells Transfer to the saliva sample collection card, and dry the collected saliva sample collection card in a pollution-free area.

[0070] Valid sample: The area where the pink area of ​​the saliva sample collection card changes to light pink or white is the valid saliva sample area.

[0071] Sample selection method: use Dubbo plastic puncher (1.0mm) for manual punching and s...

Embodiment 3

[0091] (1) Selected sample: the blood sample of subject 2.

[0092] (2) Sample type: blood sample.

[0093] (3) DNA extraction: DNA extraction is performed on the blood sample using a nucleic acid extraction instrument.

[0094] (4) System configuration

[0095]According to the instructions, prepare the primer Primer Mix and PCR reaction solution on ice according to the ratio of the instructions to configure the reaction system. After vortexing and mixing, centrifuge with a centrifuge, and then use a pipette tip to mix and distribute.

[0096] (5) Adding samples: According to the instructions, take a certain amount of extracted DNA samples with a pipette and add them to the reaction system.

[0097] (6) Amplification program

[0098] The amplification program on the PCR instrument is shown in Table 2.

[0099] (7) The amplified product is detected on the 3500DX genetic analyzer

[0100] The sample loading mixture is composed of deionized formamide and molecular weight int...

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Abstract

The invention discloses a kit and a method for detecting human pressure sensitivity genotypes. The kit and the method adopt multiple PCR amplification and electrophoresis methods to analyze and identify allele polymorphism (SNP) of five genes: MAOA, PPP1R1B, COX10, CADM2 and COMT. The method comprises the following steps: a) collecting oral cast-off cells of a testee and storing the oral cast-off cells in a collection card or performing DNA nucleic acid extraction on a blood sample; b) adding a saliva collection card sample or an extracted DNA sample of the testee into a reaction system, and performing PCR amplification; c) running an amplification program; d) performing electrophoresis analysis on an amplification product, and performing interpretation according to a peak pattern graph. Results are analyzed and interpreted. The kit and the method can synchronously detect the SNP of a plurality of related genes of the testee, and simplicity, high efficiency and specificity of detection are realized. Reference is provided for the pressure resistance of the testee through the analysis.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a kit and method for detecting human pressure sensitivity genotype. Background technique [0002] The pace of life in modern society is fast, life requirements are high, and work load is heavy. Many office workers are generally facing tremendous psychological pressure. Psychological stress will then affect daily work, study and physical health. Stress is emotionally manifested as irritability, impatience, worry, tension, indifference, anxiety, collapse, etc. Anger is one of the basic emotions inherent in animals and humans of all races. Anger is often combined with aggressive behavior and is a common manifestation of stress. Molecular genetics has proved that about 50% of the variation in stress-related behaviors is caused by genes, and studies on those gene loci that affect stress emotions have begun. These genes include monoamine oxidase A (Monoamine oxidase A, MAOA), DARPP-32...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6888C12Q1/6858C12Q2600/156C12Q2600/16C12Q2600/166C12Q2531/113C12Q2537/143C12Q2545/101C12Q2565/125
Inventor 王伟建徐智孔咪咪余丁吴勇
Owner NINGBO HEALTH GENE TECHNOLOGIES CO LTD
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