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Folate metabolism related gene MTHFR heritable variation detection kit and application thereof

A detection kit, a technology for folic acid metabolism, applied in the fields of biotechnology and clinical molecular diagnosis, can solve the problems of decreased enzyme activity and thermal stability, increased risk of neonatal birth defects or spontaneous abortion, decreased activity, etc., and achieve production costs. Low, high specificity for preventing birth defects and stroke in hypertensive patients

Inactive Publication Date: 2016-07-27
SHANGHAI HEDUN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Reduced MTHFR enzyme activity caused by mutations in the MTHFR gene can inhibit the conversion of homocysteine ​​to methionine, resulting in hypofolate and hyperhomocysteinemia, which increase the risk of birth defects or spontaneous miscarriage in newborns and other risks, as well as the occurrence of cancer, cardiovascular disease and other diseases in adults
[0007] rs1801133 (C677T) is a C>T missense mutation, which causes the 222nd amino acid of the protein encoded by the MTHFR gene to change from Ala to Val, resulting in a decrease in enzyme activity and thermal stability

Method used

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  • Folate metabolism related gene MTHFR heritable variation detection kit and application thereof
  • Folate metabolism related gene MTHFR heritable variation detection kit and application thereof
  • Folate metabolism related gene MTHFR heritable variation detection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] 1. Human Genomic DNA Extraction

[0028] Suitable samples for this kit include but are not limited to sample types such as whole blood, saliva, or oral swab. Genomic DNA extraction uses commercial kits to extract and purify DNA in tissues.

[0029] 2. Design primers according to the characteristics of this method, and the processing method is as shown in Table 1.

[0030] Table 1. Primer sequences

[0031]

[0032] 3. The fluorescent quantitative PCR reaction system is shown in Table 2:

[0033] Table 2. PCR reaction system

[0034] Composition

volume

2x HRM PreMix

25 μL

Forward primer (10μM)

0.6μL

Reverse primer (10μM)

0.6μL

Genomic DNA (20-100ng / μL)

1μL

wxya 2 o

22.8μL

[0035] 4. The PCR reaction program is shown in Table 3:

[0036] Table 3. PCR reaction program

[0037]

[0038] 5. Result analysis

[0039] refer to figure 1 According to the position of the fluorescent quantit...

Embodiment 2

[0044]A total of 20,000 pregnant women were included and randomly divided into two groups with 10,000 cases in each group. One of the groups was the observation group, taking different doses of folic acid according to the rs1801133 genotype. The other group is the control group, taking 400mg folic acid / day.

[0045] 1. Genotyping of rs1801133 in the observation group

[0046] rs1801133 was genotyped according to steps 1, 2, 3, 4 and 5 described in Example 1.

[0047] 2. The dosage of folic acid in the observation group

[0048] According to the genotype of rs1801133, the dosage of folic acid is: the recommended dosage of CC genotype is 400mg / day; the recommended dosage of CT genotype is 600mg / day; the recommended dosage of TT genotype is 800mg / day (see Table 4 in Example 1 ). The time to take folic acid is from 3 months before pregnancy to 13 weeks in the first trimester.

[0049] 3. The dosage of folic acid in the control group

[0050] The dosage of folic acid is unifo...

Embodiment 3

[0055] A total of 18317 hypertensive patients were included and randomly divided into 2 groups. One of them was the observation group (9159 cases), who received antihypertensive and / or lipid-lowering treatment and took different doses of folic acid according to the rs1801133 genotype. The other group was the control group (9158 cases), who received antihypertensive and / or lipid-lowering treatment without taking folic acid.

[0056] 1. Genotyping of rs1801133 in the observation group

[0057] rs1801133 was genotyped according to steps 1, 2, 3, 4 and 5 described in Example 1.

[0058] 2. The dosage of folic acid in the observation group

[0059] According to the genotype of rs1801133, the dosage of folic acid is: the recommended dosage of CC genotype is 400mg / day; the recommended dosage of CT genotype is 700mg / day; the recommended dosage of TT genotype is 1000mg / day (see Table 4 in Example 1 ).

[0060] 3. Follow-up results

[0061] During the 4.5 years of follow-up, there ...

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Abstract

The invention discloses a folate metabolism related gene MTHFR heritable variation detection kit.The kit is used for identifying rs1801133 genotypes and contains that 1, forward and reverse primers: the forward primer is a nucleotide strand or its complementary strand of a sequence shown by SEQ ID NO.1, and the reverse primer is a nucleotide strand or its complementary strand of a sequence shown by SEQ ID NO.2; 2, a PCR reaction: 2 x HRM PreMix.In addition, the invention further discloses application of the kit.The detection kit adopts polymerase chain reaction, a reaction system contains specific forward and reverse primers, and the genotypes are judged through real-time fluorescence quantification detection.The folate metabolism related gene MTHFR heritable variation detection kit has the advantages of being low in cost, good in specificity, high in sensitivity, simple and quick to operate and accurate in classification, making results easily interpreted and the like, can be used for clinic detection of folate metabolism related gene MTHFR heritable variation and assists a doctor to select an appropriate dose of folic acid.

Description

technical field [0001] The invention belongs to the fields of biotechnology and clinical molecular diagnosis, and specifically relates to a kit for detecting genetic variation of folic acid metabolism-related gene MTHFR; in addition, the invention also relates to the application of the kit. Background technique [0002] Numerous studies in recent years have confirmed that folic acid deficiency is the main cause of birth defects. Folic acid belongs to the B vitamins. The human body cannot synthesize folic acid and must rely on exogenous supplies. Folic acid participates in the formation and metabolism of many important compounds such as amino acid conversion and DNA synthesis in the form of tetrahydrofolate in the human body. It is a necessary substance for cell growth and tissue repair, and is an indispensable nutrient for embryonic development. Studies by Wilson et al. and Moslev et al. have shown that folic acid deficiency in pregnant women is an important risk factor for...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6883C12Q2600/106C12Q2600/156
Inventor 朱明李建新
Owner SHANGHAI HEDUN BIOTECH CO LTD
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