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SNP loci associated with obesity and/or hypertriglyceridemia in Chinese children and their application

A high triacylglycerol, site technology, applied in the field of biotechnology and medical detection

Active Publication Date: 2022-03-01
首都儿科研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 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. Open up new avenues for drug screening, efficacy evaluation and targeted therapy

Method used

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  • SNP loci associated with obesity and/or hypertriglyceridemia in Chinese children and their application
  • SNP loci associated with obesity and/or hypertriglyceridemia in Chinese children and their application
  • SNP loci associated with obesity and/or hypertriglyceridemia in Chinese children and their application

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Experimental program
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Effect test

Embodiment 1

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

[0052] From April 2004 to December 2014, the inventor collected a large number of blood samples of obese cases and control children in elementary and middle schools in Beijing and Tonghua. After sorting out the sample data, the inventor selected the samples that meet the following criteria for whole genome Experimental samples for 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]Human genomic DNA was extracted from the blood samples of obese children and normal weight children who met the above conditions 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);

[...

Embodiment 3

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

[0071] Whole-genome exome sequencing was performed on blood sample DNA of 96 obese children and 96 normal-weight children in Example 2, and candidate gene loci were obtained by comparing the exome sequencing data of the two groups of children. 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 pol...

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Abstract

The invention discloses the application of SNP sites in the preparation of early detection products for Chinese children's obesity and / or hypertriglyceridemia. The SNP sites are rs189326455, specifically chr1:233463936, c.162G>C, p.Glu54Asp . The present invention studies the genetic locus rs189326455 related to obesity as an early biomarker of obesity and hypertriglyceridemia, in order to further study the genetic molecular mechanism of obesity and hypertriglyceridemia, and to explore the early prevention and treatment of obesity and hypertriglyceridemia The drug target provides a new direction.

Description

technical field [0001] The present invention relates to the technical fields of biotechnology and medical detection, in particular to SNP sites related to obesity and / or hypertriglyceridemia in Chinese children and their application, the SNP site is rs189326455, specifically chr1:233463936, c. 162G>C, p.Glu54Asp. Background technique [0002] Obesity is a common nutritional metabolic disease. With the improvement of material living conditions in modern society, changes in diet structure, accelerated pace of life, and factors such as mental stress and lack of exercise, the number of overweight and obese people is increasing day by day . Existing evidence shows that obesity is closely related to hypertension, diabetes, dyslipidemia, and metabolic syndrome, and brings about a high risk of cardiovascular 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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/156
Inventor 米杰张美仙吴建新赵小元
Owner 首都儿科研究所
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