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Combination of mutation sites for detection of type 2 diabetes susceptibility genes, detection primers and applications

A type 2 diabetes and mutation site technology, applied in the field of molecular biology, can solve problems such as high cost, long detection time, and low accuracy

Active Publication Date: 2019-03-12
BEIJING COMPUTING CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, blood routine testing is mostly used for the detection of type 2 diabetes. This method can only be detected in patients who are already suffering from the disease, and cannot give warning and guidance to potential patients with diabetes.
[0004] According to existing research reports, no less than 17 genes are known to be related to type 2 diabetes, among which there are 25 mutation sites potentially related to type 2 diabetes, but there is no single site that can determine diabetes. There is also no system or process for association analysis of all loci
If all known mutation sites are detected, there will be problems of high cost, long detection time and low accuracy

Method used

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  • Combination of mutation sites for detection of type 2 diabetes susceptibility genes, detection primers and applications
  • Combination of mutation sites for detection of type 2 diabetes susceptibility genes, detection primers and applications
  • Combination of mutation sites for detection of type 2 diabetes susceptibility genes, detection primers and applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] This example is used to illustrate the method of using the primer combination described in the present invention to detect the mutation site described in the present invention, and the specific steps are as follows:

[0034] 1. Peripheral blood extraction

[0035] Whole blood was frozen in EDTA anticoagulant tubes at -80°C, and DNA was extracted using the QIAAmpgDNABlood Minikit.

[0036] 1) Take 200 μL blood sample with added anticoagulant to a 1.5 mL centrifuge tube, then add 40 μL proteinase K solution,

[0037] Then add 200μL AL Buffer, and immediately vortex and oscillate thoroughly for 15 seconds;

[0038] 2) Incubate at 56°C for 30 minutes;

[0039] 3) Take out the cracked specimen from the oven and shake it shortly;

[0040] 4) Add 200 μL of absolute ethanol, immediately moisten and vortex to fully mix, and shake briefly;

[0041] 5) Add the mixture of the previous step (including possible precipitation) into an adsorption column (the adsorption column is pl...

Embodiment 2

[0094] In this example, using the method described in Example 1, 1054 diabetic patients were tested for genes, and 1028 were determined to be type 2 diabetic patients, with a detection accuracy rate of 97.5%. Further, for 512 Asian healthy persons data from the 1000G (Thousand Genomes Project) public database, the genotypes of the 21 mutation sites described in the present invention were analyzed, and the formula in Example 1 was used to calculate according to the analysis results, and judged 451 were non-type 2 diabetes patients, and the detection accuracy rate was 88.1%. The comprehensive detection accuracy rate is 92.8%,

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Abstract

The invention relates to molecular biology, in particular to a mutation site combination and a primer for detecting susceptible genes of type 2 diabetes as well as an application of the combination and the primer. The mutation site combination comprises 21 mutation sites from KCNQ1, C2CD4A, TCF7L2, SLC30A8, CDKAL1, CDKN2A / 2B, KCNJ11, PPAR2, IGF2BP2, FTO, JAZF1, GRK5, RASGRP1, HHEX, PTPRD and SSR genes. Experimental research finds that the selected mutation site combination can effectively judge whether the sample is type 2 diabetes when used for detecting mutation site genes. The best detection combination is screened out from 21 SNP sites disclosed in the prior art, and the detection accuracy rate is increased to 92% or above on the basis of cost control.

Description

technical field [0001] The invention relates to molecular biology, in particular to detection of mutation site combination of susceptibility gene for type 2 diabetes, detection primers and application thereof. Background technique [0002] Type 2 diabetes mellitus (T2DM) is a complex disease caused by the interaction of genetic and environmental factors. The familial aggregation of T2DM, the high concordance of identical twins, and the high incidence in certain ethnic groups all strongly suggest the role of genetic factors in its pathogenesis. Related studies estimate that 30% to 70% of the risk of T2DM patients can be attributed to genetic susceptibility. At the same time, T2DM has genetic heterogeneity, and its phenotype is jointly controlled by polygenic genetic background and complex environmental factors. Specifically, each patient may have a different combination of susceptibility genes, and non-patient populations can also aggregate various genetic and environmental...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 李春瑞陈禹保裴智勇刘满姣卜祥霞邢玉华
Owner BEIJING COMPUTING CENT
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