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Method and kit for detecting human alcohol metabolizing capacity gene mutation sites

A technology of metabolic ability and detection method, which is applied in the field of detection method and kits of gene mutation sites of human alcohol metabolism ability, can solve the problems of difficult to meet the requirements of clinical inspection, difficult to obtain accurate detection results, and long detection cycle, etc. The results are clear and easy to understand, the experiment automation is high, and the personnel operation time is short

Inactive Publication Date: 2018-12-11
苏州道尔盾基因科技有限公司
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Problems solved by technology

[0003] The current methods for determining ADH1B and ALDH2 genotypes mainly use methods such as polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), manual or automatic sequencing, and sequence-specific primer PCR. The operation is cumbersome and the detection cycle is long. The test results are not easy to be accurate, and it is difficult to meet the requirements of clinical testing

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  • Method and kit for detecting human alcohol metabolizing capacity gene mutation sites
  • Method and kit for detecting human alcohol metabolizing capacity gene mutation sites
  • Method and kit for detecting human alcohol metabolizing capacity gene mutation sites

Examples

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Embodiment 1

[0076] Example 1: Detection of human alcohol dehydrogenase ADH1B gene rs1229984 site and acetaldehyde dehydrogenase ALDH2 gene rs671 site by nucleic acid mass spectrometry detection method

[0077] 1. Extraction of Human Genomic DNA

[0078] (1) Genomic DNA extraction is performed on the patient's blood sample in a biological safety cabinet. Extraction experiments were performed using Blood&CellCulture DNA Mini Kit (Qiagen Cat No: 13323).

[0079] (2) Primer design

[0080] SNP site primers were designed for the rs1229984 site of the human alcohol dehydrogenase ADH1B gene and the rs671 site of the acetaldehyde dehydrogenase ALDH2 gene, and specific PCR primers and single-base extension primers were designed for this SNP. The designed PCR Primer sequences are shown in Table 1.

[0081] Table 1

[0082]

[0083] (3) Specific PCR reaction

[0084] Use specific PCR technology to amplify the target fragment containing the rs1229984 site of the alcohol dehydrogenase ADH1B ge...

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Abstract

The invention relates to a method and kit for detecting human alcohol metabolizing capacity gene mutation sites. The detection method comprises the following steps: designing specific primers by aiming at an alcohol dehydrogenase ADH1B gene rs1229984 site and an acetaldehyde dehydrogenase ALDH2 gene rs671 site, and performing specific polymerase chain reaction (PCR) amplification so as to obtain target fragments; and then performing enzyme disgestion, single base extension and desalting purification treatment, and then detecting analytic sequences by use of a nucleic acid velocitron. The method for detecting the human alcohol metabolizing capacity gene mutation sites is high in detection accuracy, high in experiment repeatability, great in flux and low in cost. The invention also providesa kit for detecting human alcohol metabolizing capacity gene mutation sites. The kit comprises a specific primer pair for amplifying an alcohol dehydrogenase ADH1B gene rs1229984 site gene and an acetaldehyde dehydrogenase ALDH2 gene rs671 site gene. The kit for detecting human alcohol metabolizing capacity gene mutation sites is capable of simplifying experimental procedures and has the advantages of short manual operation time, low difficulty and high experiment automation degree.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a detection method and a kit for a mutation site of a human alcohol metabolism ability gene. Background technique [0002] The chemical name of alcohol is ethanol. After drinking, ethanol is absorbed into the blood through the stomach. 90%-95% of ethanol entering the body is metabolized through the liver, and the rest is excreted with urine, sweat and breath. Ethanol mainly undergoes two steps of metabolic reactions in the liver: oxidation from ethanol to acetaldehyde; reoxidation from acetaldehyde to acetic acid. These two steps of metabolic reactions are all carried out under the catalysis of enzymes. The former enzyme is called alcohol dehydrogenase (ADH), and the latter enzyme is called acetaldehyde dehydrogenase (ALDH). Studies by genetic scientists have shown that alcohol dehydrogenase is mainly encoded by the ADH1B gene, and acetaldehyde dehydrogenase is mai...

Claims

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Application Information

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IPC IPC(8): C12Q1/6806C12Q1/6872C12Q1/6883
CPCC12Q1/6806C12Q1/6872C12Q1/6883C12Q2600/156C12Q2531/113C12Q2521/301C12Q2533/101C12Q2565/627
Inventor 王文忠胡军田军龙陈苏平卜云璇
Owner 苏州道尔盾基因科技有限公司
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