Human MTHFR and MTRR gene detection kit and application thereof

A detection kit and gene detection technology, applied in the biological field, can solve the problems of prolonged diagnosis time, low sensitivity, cumbersome and time-consuming operation, etc., and achieve the effects of strong repeatability, simple operation, and rapid and objective detection results.

Active Publication Date: 2018-07-06
JIANGSU BIOPERFECTUS TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to complete the detection of the genotypes of the three loci of the MTHFR and MTRR genes, a fluorescent PCR reaction is required, which will undoubtedly consume a lot of manpower...

Method used

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  • Human MTHFR and MTRR gene detection kit and application thereof
  • Human MTHFR and MTRR gene detection kit and application thereof
  • Human MTHFR and MTRR gene detection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Assembly of human MTHFR and MTRR gene detection kits

[0044] The kit includes the following components:

[0045] (1) Nucleic acid amplification reaction solution:

[0046] The nucleic acid amplification reaction solution is composed of RNase-free water, 10×PCR buffer, 25mM Mg 2+ , 10mM dNTPs composition (Table 1).

[0047] The composition of table 1 nucleic acid amplification reaction solution

[0048] Reagent name

Adding volume (μL) / 50 servings

PCR buffer

100

MgCl 2

3

100mM dATP

2.5

100mM dGTP

2.5

100mM dCTP

2.5

100mM dUTP

1.25

100mM dTTP

1.25

RNase-free water

337

total capacity

450

[0049] (2) Enzyme mixture

[0050] The enzyme mixture includes DNA polymerase, enzyme diluent and UDG enzyme. DNA polymerase plays a very important role in PCR amplification, including the efficiency and specificity of reverse amplification. In order to achieve bett...

Embodiment 2

[0079] Adopt the test kit of the embodiment of the present invention 1 to the detection test of clinical sample

[0080] 1. Sample processing (whole blood sample extraction)

[0081] 200 μL of whole blood samples were taken, and Qiagen Whole Blood Genomic DNA Extraction Kit (QIAamp DNA BloodMini Kit, Cat No.: 51104) was used to extract according to the kit instructions to obtain sample DNA.

[0082] 2. Reagent preparation

[0083] The kit prepared in Example 1 was used to prepare the reaction system (Table 7):

[0084] Table 7 Reaction system configuration table

[0085] reaction system

Dosage (μL) / 1 person

Nucleic Acid Amplification Reaction Solution

9

Enzyme Mix

1

Detection solution

8

sample DNA

2

total capacity

20

[0086] 3. PCR amplification program

[0087] Perform RT-PCR amplification according to the following procedure: amplification (37°C for 5min; 95°C for 15min; 95°C for 10s, 55°C for 40s...

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PUM

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Abstract

The invention discloses a human MTHFR and MTRR gene detection kit and an application of the human MTHFR and MTRR gene detection kit. The kit comprises the following components: a MTHFR 677/MTHFR 1298/MTRR 66 triplet detection liquid and the following optional components of a nucleic acid amplification reaction liquid, an enzyme mixed solution, a positive control and a blank control. The disclosedhuman MTHFR and MTRR gene detection kit can detect genotypes of a 677 locus and a 1298 locus of a MTHFR gene and a lotus 66 of a MTRR gene at the same time in a same reaction tube. In addition, the kit further has the advantages of strong specificity, high sensitivity, convenient operation, strong repeatability, rapid and objective detection result and the like. The provided in vitro detection ofthe human MTHFR and MTRR genes provides an effective technical means.

Description

technical field [0001] The invention relates to an in vitro diagnostic kit for human genome DNA genotype detection, in particular to a MTHFR and MTRR gene detection kit and its application, and belongs to the field of biotechnology. Background technique [0002] Folate is an important methyl donor in the diet, involved in DNA methylation and DNA synthesis pathways. The lack of folic acid will cause uridine in human DNA to be inserted incorrectly and break chromosomes, which will lead to the instability of the genome and increase the probability of mutation. Abnormalities in the folate metabolism pathway lead to hyperhomocysteine ​​sepsis. Hyperhomocysteine ​​sepsis is a risk factor for several diseases and leads to various increased risks such as vascular and neurological diseases, pregnancy complications, male infertility, birth defects, diabetes, kidney disease, osteoporosis , neuropsychiatric disorders and cancer. [0003] The MTHFR (5,10-methylenetetrahydrofolate redu...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6886
CPCC12Q1/6883C12Q1/6886C12Q2600/156
Inventor 巫益鸣胡艳民徐加发刘中华王国强
Owner JIANGSU BIOPERFECTUS TECH CO LTD
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