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Method and kit for detecting genes related to folic acid metabolism

A folic acid metabolism and kit technology, which can be used in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of reduced enzyme activity and affect folic acid metabolism, and achieve high accuracy, low cost, and simple operation. Effect

Active Publication Date: 2020-01-17
天津华大基因科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Mutations in the genes of these two enzymes lead to reduced enzyme activity, which will significantly affect the body's ability to metabolize folic acid

Method used

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  • Method and kit for detecting genes related to folic acid metabolism
  • Method and kit for detecting genes related to folic acid metabolism
  • Method and kit for detecting genes related to folic acid metabolism

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Collect and select 66A of MTRR gene> G site and 677C of the MTHFR gene> T site and 1298A> There are 3 sites related to folate metabolism in C site. Hereinafter, the first primer is also called an amplification primer, and the second primer is also called an extension primer. It is not emphasized that the primer of the first or second primer may include the first primer and / or the second primer.

[0037] Design and optimization of amplification primers and extension primers for the 7 sites, primer design can be performed after determining the upstream and downstream sequences of the target site, so that there is no serious dimer, mismatch and hairpin between the amplification primers or extension primers , The length of the amplification primer is about 30 bases, the amplification primer has a tag sequence of 10 bases acgttggatg at the 5'end, and the length of the extension primer is about 17-28 bases, allowing extension of the 5'end of the primer 1 -5 bases are not complet...

Embodiment 2

[0040] 1) DNA extraction

[0041] Collected 18 human whole blood samples, and extracted DNA using chelex-100 (resin method).

[0042] 2) PCR amplification

[0043] Through this step of PCR amplification, a DNA fragment containing the target site is obtained. Refer to Table 5 for the PCR amplification reaction system. Among them, all reagents were purchased from Sequenom, USA, and the PCR instrument was PCR System 9700Dual 384-Well Sample Block Module.

[0044] table 5

[0045]

[0046] The PCR reaction conditions were 94°C for 2 minutes; 94°C denaturation for 20 seconds, 56 annealing for 30 seconds, 72 extension for 1 minute, a total of 45 cycles of amplification; and finally 72°C for 5 minutes extension.

[0047] In PCR, the positive control used is normal human genomic DNA with known sequence, and the negative control is sterile double distilled water.

[0048] 3) SAP processing

[0049] The amplified product in the previous step was treated with SAP enzyme (shrimp alkaline phosphata...

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Abstract

The invention discloses a method for detecting folate metabolism-related gene. The method comprises the following steps: subject nucleic acid is obtained, the nucleic acid comprises DNA fragment; at least two SNP bits genotype are detected, MTRR gene c.66A>G; MTHFR gene c.667C>T; and MTHFR gene c.1298A>C; a first primer amplification comprise the DNA fragment of the SNP bit, the amplification product is obtained, a second primer is combined with the amplification product, a base is extended, an extension product is obtained, according to the molecular weight difference of the extension product and the second primer, the base type of the SNP bit is determined. The invention also discloses a kit for detecting folate metabolism capability gene. By using the method and / or the kit, the detecting folate metabolism-related gene can be typed with high flux, low cost, high accuracy and short time consumption, and the typing result can help the subject for supplying folic acid.

Description

Technical field [0001] The present invention relates to the field of biological detection. Specifically, the present invention relates to a method for detecting genes related to folate metabolism and a kit. Background technique [0002] Folic acid (pteroyl monoglutamic acid) refers to a class of equivalent vitamins with related biological activities, which is formed by the combination of pteridine, p-aminobenzoic acid and one or more glutamic acids, namely, a-amino-4 -Hydroxypteridine is connected to p-aminobenzoic acid, and then connected to glutamic acid by -NH-CO- bond. Its different coenzyme forms can act as donors or carriers of one carbon unit, and are mainly involved in the following Class reactions: ① synthesis of methionine; ② synthesis of purine and pyrimidine; ③ mutual conversion of serine and glycine; ④ degradation of histidine. A large number of scientific studies have shown that absolute or relative lack of folic acid intake is the direct cause of hyperhomocysteine...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883
Inventor 王晓楠张俊青李智丽杨玲
Owner 天津华大基因科技有限公司
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