Method for detecting folate metabolism-related gene and kit thereof

A technology of folic acid metabolism and genetics, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of enzyme activity reduction, affecting folic acid metabolism ability, etc., and achieve the effect of high accuracy, low cost and simple operation

Active Publication Date: 2016-10-05
天津华大基因科技有限公司 +1
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

Mutations in the genes of these two enzymes lead to reduced enzyme activi...

Method used

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  • Method for detecting folate metabolism-related gene and kit thereof
  • Method for detecting folate metabolism-related gene and kit thereof
  • Method for detecting folate metabolism-related gene and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The 66A>G site of the MTRR gene and the 677C>T site and 1298A>C site of the MTHFR gene were collected and selected to be 3 sites related to folic acid metabolism ability. Hereinafter, the first primer is also referred to as an amplification primer, and the second primer is also referred to as an extension primer, and primers that do not emphasize the first or second primer may include the first primer and / or the second primer.

[0037] Design and optimize amplification primers and extension primers for the 7 sites, and design primers after determining the upstream and downstream sequences of the target sites, so that there are no serious dimers, mismatches, and hairpins between amplification primers or extension primers , so that 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 the extension primer 5' end 1 -5 b...

Embodiment 2

[0040] 1) DNA extraction

[0041] Eighteen human whole blood samples were collected and DNA was extracted using chelex-100 (resin method).

[0042] 2) PCR amplification

[0043] Through PCR amplification in this step, a DNA fragment containing the target site is obtained. See Table 5 for the PCR amplification reaction system. Among them, all reagents were purchased from Sequenom Company in the United States, and the PCR instrument was PCR System 9700Dual384-Well Sample Block Module.

[0044] table 5

[0045]

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

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

[0048] 3) SAP processing

[0049] The amplified product of the p...

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

Claims

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Application Information

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IPC IPC(8): C12Q1/68
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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