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One-tube ALDH2 genotyping kit and detection method thereof

A genotyping and kit technology, applied in the field of medical testing, can solve problems such as cumbersome steps, difficult census methods, and high costs, and achieve the effects of low cost, saving testing costs, and avoiding contamination risks

Active Publication Date: 2020-07-14
天筛(上海)科技有限公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, all these methods must extract the genomic DNA of the sample, making the steps cumbersome and expensive, and it is difficult to become a general survey method

Method used

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  • One-tube ALDH2 genotyping kit and detection method thereof
  • One-tube ALDH2 genotyping kit and detection method thereof
  • One-tube ALDH2 genotyping kit and detection method thereof

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Embodiment Construction

[0033] Aldehyde dehydrogenase 2 (ALDH2) is one of the members of the NAD+-dependent aldehyde dehydrogenase superfamily. At present, 19 members of the aldehyde dehydrogenase family have been found. Aldehyde dehydrogenase 2 is located in the mitochondria. An important aldehyde oxidase, which can oxidize acetaldehyde, an intermediate product of ethanol metabolism, to acetic acid. At the same time, acetaldehyde dehydrogenase 2 can decompose acetaldehyde metabolite 4-hydroxynonenal, and reduce the oxidative damage of acetaldehyde and its metabolites to cells. It is a tetrameric enzyme that is abundantly expressed in tissues such as liver, kidney and lung, and also expressed in heart and brain tissues where mitochondrial function is active. Recent studies have shown that the activation of this enzyme has a significant protective effect on myocardial cell death caused by myocardial ischemia-reperfusion. Aldehyde dehydrogenase 2 gene has polymorphism, and the 487 amino acid residues ...

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Abstract

The invention discloses a simple and economical one-tube ALDH2 genotyping kit. The kit comprises the following components: a direct-amplification dye method qPCR premixed solution containing three primer groups, a positive plasmid control and a negative control. The kit is characterized in that the three primer groups are as follows: an ALDH2-F upstream primer is shown in SEQ ID NO: 1, an ALDH2-Rdownstream primer is shown in SEQ ID NO: 2, and an ALDH2-MR mutant downstream primer is shown in SEQ ID NO: 3. The kit disclosed by the invention adopts one-tube qPCR reaction, and an extraction stepof a sample is omitted, so that the test cost is greatly saved, and the pollution risk in the extraction process is avoided. The kit is accurate, rapid and low in cost, is particularly suitable for risk analysis of genetic factors such as drinking behavior, alcoholic cirrhosis and liver cancer, and is also a reference method for clinical nitroglycerin guiding medication.

Description

technical field [0001] The invention belongs to the field of medical detection, and in particular relates to a simple and economical one-tube type ALDH2 genotyping kit and a detection method thereof. Background technique [0002] The full name of ALDH2 is aldehyde dehydrogenase 2 family (mitochondrial), and the Chinese name is aldehyde dehydrogenase 2. The ALDH2 gene is located at 12q24.2 on chromosome 12, and its main polymorphism is rs671, which is G1510A located in exon 12. A single base mutation from G to A occurs at this position, so that Glu504 is mutated to Lys504. This site is related to the spatial structure of the ALDH2 protein, thus affecting the activity of the enzyme. The normal allele is denoted as ALDH2*1, and the allele with a single base mutation is denoted as ALDH2*2. Glu504 homozygous (ALDH2*1 / 1) showed normal enzymatic activity; Glu504 / Lys504 heterozygous (ALDH2*1 / 2) had only about 6% of its normal activity; Aldehyde degradation is essentially free of ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/686C12N15/11
CPCC12Q1/6883C12Q1/686C12Q2600/156C12Q2600/106C12Q2563/107C12Q2545/114C12Q2527/107
Inventor 王继国陈普
Owner 天筛(上海)科技有限公司
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