Method for predicting risk of non-alcoholic fatty liver disease by gene polymorphism

A gene polymorphism, non-alcoholic technology, used in the field of gene polymorphism to predict the risk of non-alcoholic fatty liver disease, can solve the problem of incomplete application, incomplete distribution of genetic single nucleotide polymorphisms in ethnic groups, Lack of data and other problems, to achieve the effect of convenient and fast methods

Inactive Publication Date: 2019-06-25
QINGDAO MUNICIPAL HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Defects and insufficiencies of existing technologies: There are few studies on NAFLD susceptibility genes in my country, and there is a lack of instructive data; in view of different regions, the distribution of genetic single nucleotide polymorphisms in ethnic groups is not completely consist...

Method used

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  • Method for predicting risk of non-alcoholic fatty liver disease by gene polymorphism
  • Method for predicting risk of non-alcoholic fatty liver disease by gene polymorphism
  • Method for predicting risk of non-alcoholic fatty liver disease by gene polymorphism

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Example 1 template DNA extraction

[0086] 1) Use whole blood genomic DNA extraction kit to extract 200 μL blood sample;

[0087] 2) Combine 200ul of buffer G and 20μL of proteinase K into a premix, add the blood sample, shake and mix well;

[0088] 3) Place it at 56°C for a certain period of time, and at the same time mix it upside down until it becomes a clear liquid;

[0089] 4) Place it at room temperature for 5 minutes, add 350ul buffer solution BD, fully invert and mix, and then centrifuge briefly;

[0090] 5) Put the adsorption column into the collection tube, add the mixed solution, centrifuge at 12000rpm for 30 seconds, remove the useless liquid, and put it back into the collection tube;

[0091] 6) Take 500ul buffer solution GDB and add it to the adsorption column, centrifuge at 12000rpm for 30 seconds, remove useless liquid, and put it back into the collection tube;

[0092] 7) Add 600ul of rinse solution PWB to the adsorption column, centrifuge at 12000rp...

Embodiment 2D

[0096] Example 2 DNA concentration and purity identification

[0097] Use ultraviolet spectrophotometer to identify, use OD260nm / OD280nm ratio to calculate DNA purity, the ratio is 1.8, and according to the concentration measured, use ddH 2 0 Dilute the DNA to 20ng / μL, and finally store it in a -80°C refrigerator until use.

Embodiment 3

[0098] Example 3 Genotyping

[0099] TM6SF2rs58542926, PNPLA3 rs738409 and TRBI rs17321515 loci were genotyped using the TaqMan probe method (Beijing Bomiao Biological Company);

[0100] 1. Primers:

[0101] TM6SF2rs58542926 1:5'-ACGTTGGATGGCACCATGGAAGGCAAATAC-3';

[0102] TM6SF2rs58542926 2:5'-ACGTTGGATGTGAAGACCTTCATGCCAGCC-3';

[0103] PNPLA3rs738409: 5'-AACTTCTCTCCTTTGCTTTCACA-3';

[0104] PNPLA3rs738409: 5'-GGAGGGATAAGGCCACTGTAGA-3';

[0105] TRBI rs17321515: 5'-ACGTTGGATGGAACAAGGACTTTCGTCCTC-3';

[0106] TRBI rs17321515: 5'-ACGTTGGATGTAGAAGTCCCCTTCCCTTAG-3';

[0107] 2. PCR amplification reaction system

[0108]

[0109] SAP product alkaline phosphatase treatment

[0110]

[0111]

[0112] EX single base extension reaction

[0113]

[0114] 3. TaqMan amplification reaction program:

[0115]

[0116] 4. TaqMan amplification reaction

[0117] Configure the reaction mix according to the reaction system, and distribute it into 384-well plates; add sam...

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PUM

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Abstract

The invention discloses a method for predicting the risk of non-alcoholic fatty liver disease by genetic polymorphism. According to the method, the risk of NAFLD in the population can be predicted. Based on the comparative analysis of genetic data of a large number of NAFLD population and healthy population in China, three polymorphic sites of TM6SF2 rs58542926, PNPLA3 rs738409 and TRBI rs17321515closely related to NAFLD susceptibility are selected, and the data are statistically analyzed to calculate the risk of NAFLD susceptibility genotype in Chinese population. The method has important guiding significance for prevention and treatment of the NAFLD, and is suitable for popularization and application.

Description

technical field [0001] The present invention relates to the field of prediction of human non-alcoholic fatty liver, more specifically, relates to a method for predicting the risk of non-alcoholic fatty liver by gene polymorphism. Background technique [0002] Non-alcoholic fatty liver disease (NAFLD) is a chronic liver disease that affects both adults and children around the world. NAFLD can progress from a benign histologic disease stage characterized by pure fat accumulation (commonly referred to as simple steatosis or nonalcoholic fatty liver disease) to a more severe histologic form characterized by hepatocellular injury, mixed inflammatory lobular infiltrates, and Characterized by nonalcoholic steatohepatitis (NASH). Similar to other chronic liver diseases, both NAFLD and NASH can lead to the progression of fibrosis in some patients, and eventually develop into liver cirrhosis and its complications. Due to the late economic development in Asia, most patients have a sh...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883
Inventor 辛永宁薛枫赵本田张暘
Owner QINGDAO MUNICIPAL HOSPITAL
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