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Single nucleotide polymorphisms associated with childhood obesity in China and its application

A fat, site-based technology for biomedical applications

Active Publication Date: 2020-04-28
首都儿科研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of the fact that there are no early biomarkers and early diagnostic kits related to obesity, if the corresponding markers can be found and the corresponding diagnostic kits can be developed, it will provide a new direction for the exploration of drug targets for the early prevention and treatment of obesity, and for its Open up new avenues for drug screening, efficacy evaluation and targeted therapy

Method used

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  • Single nucleotide polymorphisms associated with childhood obesity in China and its application
  • Single nucleotide polymorphisms associated with childhood obesity in China and its application
  • Single nucleotide polymorphisms associated with childhood obesity in China and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0051] The collection of embodiment 1 sample and the arrangement of sample data

[0052] The inventor collected a large number of blood samples of obese case-control children in primary and secondary schools in Beijing and Tonghua from April 2004 to December 2014. After sorting out the sample data, the inventor selected 6334 samples that met the following criteria for Experimental samples for whole genome exome sequencing and single SNP Sequenom MassARRAY genotyping:

[0053] 1. Obese cases clearly diagnosed by the body mass index (BMI) classification criteria recommended by the International Obesity Working Group (IOTF);

[0054] 2. Han children, aged 6-18;

[0055] 3. The control included normal weight and wasted children, excluding overweight children.

[0056] The demographic data and clinical data of these samples were collected systematically.

Embodiment 2

[0057] Example 2 Peripheral Blood DNA Extraction

[0058] In all the above-mentioned eligible samples, human genomic DNA was extracted using a spin column method (DNA mini-extraction kit, QIAGEN, Germany). The specific steps are:

[0059] 1. Add 20 μL proteinase K to a 1.5 mL centrifuge tube;

[0060] 2. Add 200 μL white blood cell mixture, if the volume is less than 200 μL, make up with phosphate buffer saline (PBS);

[0061]3. Add 200 μL buffer AL (lysate), vortex for 15 seconds, and centrifuge quickly;

[0062] 4. Water bath at 56°C for 10 minutes;

[0063] 5. Centrifuge quickly to avoid leaving solution on the tube cap;

[0064] 6. Add 200 μL absolute ethanol, vortex for 15 seconds, and centrifuge quickly;

[0065] 7. Transfer all the solutions to the QIAamp spin column, centrifuge at 10000rpm for 2 minutes, discard the collection tube, and transfer the spin column to a new collection tube (provided by the kit);

[0066] 8. Add 500 μL buffer AW1 (washing solution 1) ...

Embodiment 3

[0070] Example 3 Whole Exome Detection of SNP in Peripheral Blood DNA

[0071] Whole-genome exome sequencing was performed in the 19 severely obese children and 15 emaciated children who met the above conditions, and the exome sequencing data of the two groups of children were compared to obtain candidate gene loci. The specific steps are:

[0072] 1. DNA quality inspection: Use a UV / visible spectrophotometer (DU800, Beckman, USA) or (NanoDrop 2000, Thermo Fisher, USA) to read the absorbance values ​​at A230nm, A260nm, and A280nm, and calculate the DNA concentration according to the formula (g / L)=A260nm×50, and DNA purity was identified by OD ratio (A260 / A280, A260 / A230). A low A260 / A280 ratio indicates protein residues, but if phenol is used during the operation, it is more likely to be phenol residues, and a high A260 / A280 ratio suggests that ribonucleic acid (RNA) may not be removed completely. A low ratio of A260 / A230 indicates residual salt or small molecular impurity p...

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Abstract

The invention relates to a single nucleotide polymorphism sites related to obesity of Chinese children and application thereof. The invention discloses a biomarker related to obesity of Chinese children. The biomarker comprises one or two SNP sites of susceptibility to obesity of Chinese children, wherein the sites comprises susceptibility SNP site (1): rs1059491, or in concrete, the chr16: 28603655, c. 704A> C, p. N235T; susceptibility SNP site (2): rs768847893, or in concrete, the chrX: 3241682, c. 2044> T, p. R682C. According to the single nucleotide polymorphism sites related to obesity of Chinese children, two susceptibility SNP sites are found, and the application prospect of the sites in the auxiliary tests of children obesity is investigated. The gene sites related with obesity are possibly the early biomarkers of obesity, and provide a new direction for the further investigation of the genetic molecular mechanism of obesity and the exploration of drug targets for the early prevention of obesity.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a single nucleotide polymorphism site related to childhood obesity in China and its application. Background technique [0002] Obesity is a common nutritional metabolic disease. In 2002, WHO listed obesity as one of the global first-level risk factors causing the burden of human disease. Preventing the prevalence of obesity is also one of the biggest public health challenges faced by countries in the world in the first 50 years of the 21st century. Family analysis and twin studies have shown that genetic factors play a very important role in the occurrence of obesity. Genetic variation in more than 40% of the genes associated with body fat in humans can make a difference. [0003] Single nucleotide polymorphism (single nucleotide polymorphism, SNP) is a type of molecular genetic marker proposed by Lander, a scholar at the Human Genome Research Center of the Massachusetts In...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/136C12Q2600/156
Inventor 米杰张美仙吴建新赵小元吴丽君程红侯冬青
Owner 首都儿科研究所
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