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Test kit for detecting folic acid metabolic pathway related genes

A pathway-related and folic acid metabolism technology, which is applied in the field of kits for detecting genes related to folic acid metabolism pathways, can solve the problems of complex operation, low detection rate, and reduced enzyme activity, and achieves overcoming high sample quality requirements and high detection rate. , to ensure the effect of accuracy

Active Publication Date: 2020-07-10
HAINAN ZHUJIAN CELL MOLECULE GENETICS INSPECTION CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The corresponding enzyme activity reduction caused by CBS gene, MTR gene, MTHFR gene and MTRR gene mutation inhibits the conversion of homocysteine ​​to methionine, resulting in hypofolate and hyperhomocysteinemia, thereby increasing Risk of spontaneous miscarriage or birth defects in pregnant women
[0005] The existing detection of genes related to folic acid metabolism pathway is mainly aimed at polymorphism detection of MTHFR gene and MTRR gene loci, and the existing detection technology is easy to operate due to its complicated operation when the number of detection sites increases. The sample quality requirements are high, the detection rate is low, the stability and efficiency of the experiment are low, and it is difficult to achieve efficient and accurate simultaneous detection of polymorphic sites in multiple key genes of folic acid metabolism pathways, which limits the use of this detection product in guiding Effective application of reference for women who are planning to become pregnant and pregnant women to take folic acid reasonably

Method used

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  • Test kit for detecting folic acid metabolic pathway related genes
  • Test kit for detecting folic acid metabolic pathway related genes
  • Test kit for detecting folic acid metabolic pathway related genes

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0038] Example 1 - Design of Primer Compositions

[0039] The present invention optimizes and selects 5 SNP site systems on MTR gene, CBS gene, MTRR gene and MTHFR gene, and designs 5 pairs of PCR primers for amplifying DNA fragments containing 5 SNP sites, and designs The composition of 5 forward extension primers next to the SNP site; at the same time, the extension primers of different lengths were designed for different SNP sites, that is, the number of poly T modified on the 5 kinds of single base extension primers and the single The concentration of base extension primers makes the extension products produced by their respective genes have different molecular weights, so that multiple SNP sites can be accurately typed in one reaction system, and can be determined efficiently and intuitively according to the moving position of the electrophoretic front The SNP sites corresponding to different extension products can directly determine the type of base incorporated accordin...

Embodiment 2

[0043] Preparation and use of embodiment 2-kit

[0044] a. The components and contents of the kit include:

[0045] (1) Multiplex PCR amplification reagents: dNTPs, TaqDNA polymerase, PCR reaction buffer, 1x GC-I buffer, Mg2+:

[0046] (2) PCR product purification reagents: AP enzyme, Exol1 enzyme, Seq buffer;

[0047] (3) Multiple single base extension reaction reagent: SNaPshot Multiplex Kit (ABI);

[0048] (4) Extension product purification reagent: SAP enzyme;

[0049] (5) Sequencing denaturation reagents: 120 Liz SIZE STANDARD internal standard, Hi-Di formamide.

[0050] b. Use of kits

[0051] 1. Sample sampling

[0052] Take a saliva sample from the oral cavity test and extract DNA to obtain 1 μl of DNA sample. 1% agarose electrophoresis is used for quality inspection and concentration estimation of the sample, and then the sample is divided into 4 equal parts according to the estimated concentration and diluted to a concentration of 0.8ng. / μl, 1ng / μl, 5ng / μl, 10...

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Abstract

The invention provides a test kit for detecting folic acid metabolic pathway related genes. The kit comprises primer compositions for detecting an MTR gene RS1805087 site, a CBS gene RS5742905 site, aMTRR gene RS1801394 site, a MTHFR gene rs1801131 site and a MTHFR gene RS1801133 site, which are related to a folic acid metabolic pathway, the sequences of the primer compositions comprise five pairs of PCR primers and five forward extension primer compositions adjacent to SNP sites, and the forward extension primer compositions are shown as SEQ ID No: 1-15. The test kit not only can realize one-time multi-gene-site detection, but also reduces the lowest detection limit to 1ng / Mul, is high in detection rate, realizes large-flux rapid genotyping of gene site, and ensures the accuracy of sample detection to be 100%.

Description

technical field [0001] The invention relates to the technical field of gene polymorphism detection, in particular to a kit for detecting genes related to folic acid metabolic pathways. Background technique [0002] Folic acid is an important nutrient required by the body. It is a water-soluble B vitamin and participates in a series of important biochemical reactions involving the transfer of one-carbon units. With the continuous advancement of medical science such as medical genetics and molecular biology, people are increasingly aware of many new important functions of folic acid, especially the lack of folic acid will lead to chromosome damage and breakage, and the frequency of sister chromosome exchange will increase. The expression of fragile sites can cause mutagenesis and even canceration in severe cases. Folic acid is an essential element for the synthesis of nucleic acid, an essential substance for cell growth and tissue repair, and an indispensable nutrient for emb...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 陈春宝苏震张子询
Owner HAINAN ZHUJIAN CELL MOLECULE GENETICS INSPECTION CENT
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