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Kit for detecting SNP sites related to homocysteine metabolism and amplification method and detection method thereof

A technology of homocysteine ​​and kits, which is applied in the field of kits for detecting SNP sites related to homocysteine ​​metabolism and its amplification and detection, can solve the problems of reduced enzyme activity and increased HCY levels, and achieves The effect of reducing operation steps, good repeatability, and avoiding false negative results

Active Publication Date: 2013-07-03
UNION STEMCELL & GENE ENG +1
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AI Technical Summary

Problems solved by technology

The mutation of related enzyme gene in the metabolism of folic acid and HCY will lead to the reduction of enzyme activity and increase the level of HCY

Method used

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  • Kit for detecting SNP sites related to homocysteine metabolism and amplification method and detection method thereof
  • Kit for detecting SNP sites related to homocysteine metabolism and amplification method and detection method thereof
  • Kit for detecting SNP sites related to homocysteine metabolism and amplification method and detection method thereof

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Embodiment Construction

[0041] The kit for detecting single nucleotide polymorphism sites of genes related to homocysteine ​​metabolism in the present invention, the kit can simultaneously detect the rs1801133 SNP site of the MTHFR gene, the rs1805087 SNP site of the MTR gene, and the rs1801394 SNP site of the MTRR gene and the CBS gene rs5742905 SNP site for amplification and typing detection, and the ACTB gene as an internal reference to detect the amplification conditions; the wild-type 2× buffer contains the wild-type forward sequence-specific primers of the above 4 SNP sites, and the common reverse sequence The primers and the upstream and downstream primers of the internal reference sequence are used to determine whether the wild-type phenotype corresponding to the SNP site is carried by the presence or absence of the DNA electrophoresis band after amplification; Sequence-specific primers, common reverse primers, and internal reference sequence upstream and downstream primers are used to determi...

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Abstract

The invention discloses a detection kit for detecting SNP sites related to homocysteine metabolism by using multiplex PCR technology and SNP sensitive molecule switch technology and an amplification method and a detection method thereof. The kit carries out genetyping on four SNP sites related to homocysteine metabolism, i.e., an rs1801133 SNP site in the MTHFR gene, an rs1805087 SNP site in the MTR gene, an rs1801394 SNP site in the MTRR gene and an rs5742905 SNP site in the CBS gene. The kit comprises a wild 2* amplification buffer, a mutant 2* amplification buffer, polymerase, a cell lysis solution and glycerin, wherein the two buffers respectively contain sequence-specific primers and internal control primers corresponding to SNP phenotypes consisting of a wild type and a mutant type, genetyping of the above-mentioned SNP sites can be finished in two multiplex PCR reactions, so homocysteine metabolic capability and demands for folic acid, VB12 and VB6 of a subject are known.

Description

technical field [0001] The present invention relates to a kit for detecting SNP sites related to homocysteine ​​metabolism by using multiplex PCR combined with molecular switch technology and its amplification method and detection method, in particular to a MTHFR gene rs1801133 for detecting homocysteine ​​metabolism The invention relates to a test kit for single nucleotide polymorphism (SNP) of SNP site, MTR gene rs1805087 SNP site, MTRR gene rs1801394 SNP site and CBS gene rs5742905 and a multiplex PCR amplification method, belonging to the field of biomedicine. Background technique [0002] Homocysteine ​​(HCY), also known as homocysteine ​​or homocysteine, is formed from S-adenosylmethionine through a two-step reaction pathway and can be converted to formazan Thionine, or converted to cysteine ​​or taurine through the transsulfur pathway. Lack of vitamins such as folic acid, VB6 or VB12 will increase the level of HCY. Supplementation of folic acid, VB6, VB12 or trimeth...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 韩俊领杜宏伟周毓玲崔丽娟
Owner UNION STEMCELL & GENE ENG
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