Method for detecting Type 2 diabetes related mononucleotide polymorphism by using DNA (deoxyribonucleic acid) microarray chip

A technology of microarray chip and diabetes, which is applied in the determination/testing of microorganisms, biochemical equipment and methods, etc.

Active Publication Date: 2015-03-04
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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  • Description
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Problems solved by technology

At present, a gold nanoparticle marker can usually only be detected for one biomolecule. Therefore, preparing a gold nanoparticle marker that can detect multiple analytes at the same time can simplify the detection method of microarray biochips. of great significance

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  • Method for detecting Type 2 diabetes related mononucleotide polymorphism by using DNA (deoxyribonucleic acid) microarray chip
  • Method for detecting Type 2 diabetes related mononucleotide polymorphism by using DNA (deoxyribonucleic acid) microarray chip
  • Method for detecting Type 2 diabetes related mononucleotide polymorphism by using DNA (deoxyribonucleic acid) microarray chip

Examples

Experimental program
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Effect test

Embodiment 1

[0110] Embodiment 1: the detection of DNA sample of simulated type Ⅱ diabetes

[0111] combine figure 1 , image 3 , Figure 4 and Table 1 illustrate this example.

[0112] Using the standard method developed by the Brown Laboratory of Stanford University in the United States, a fixed concentration (30 μmol / L) of oligonucleotide probe (P 1 -P 6 ) were spotted on the substrate to prepare a DNA microarray chip. The chip was blocked by using PBS (PH=7.4, 50 mmol / L phosphate buffer + 0.15 mol / L sodium chloride) and 0.1 mol / L ethanolamine. and in turn with the oligonucleotide target T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 1A , T 2A , T 3A , T 4A , T 5A and T 6A The concentrations are: 0.005nM, 0.01nM, 0.05nM, 0.1nM, 0.5nM, 1nM, 5nM, 10nM, 50nM, 100nM, 500nM and 1000nM mixture, T 1A -T 6A The percentages in the mixture were: 0, 0.2, 0.5, 1, 2, 5, 10, 20, 50, 70, 90 and 100%, and 3nM oligonucleotide-modified gold nanoparticles (SP n GNPs), each reacted at 30°C for 1...

Embodiment 2

[0115] Embodiment 2: the actual sample detection of type Ⅱ diabetes

[0116] combine figure 1 , Figure 5 and Table 1 illustrate this example.

[0117] The blood of 15 patients with type Ⅱ diabetes was collected, and PCR amplification was performed for the rs10811661 locus. Using the standard method developed by the Brown Laboratory of Stanford University in the United States, a fixed concentration (30 μmol / L) of oligonucleotide probe (P n ) were spotted on the substrate to prepare a DNA microarray chip. The chip was blocked by using PBS (PH=7.4, 50 mmol / L phosphate buffer + 0.15 mol / L sodium chloride) and 0.1 mol / L ethanolamine. And in turn with different concentrations (0.5nM, 1nM, 5nM and 10nM) of type Ⅱ diabetes patients blood PCR products, and 3nM oligonucleotide modified gold nanoparticles (SP nGNPs), each reacted at 30°C for 1 h. After further reacting with 1mL silver enhancement solution for 8 minutes, detection was performed using a resonance light scattering mi...

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Abstract

The invention relates to a method for detecting Type 2 diabetes related mononucleotide polymorphism by using a DNA (deoxyribonucleic acid) microarray chip, belonging to the technical field of DNA microarray chips. By using 6-oligonucleotide-modified gold nanoparticles as the markers in combination with 6 oligonucleotide probes for Type 2 diabetes detection, a microarray chip inspection method capable of simultaneously detecting 6 Type 2 diabetes related mutant sites is developed, thereby implementing the detection on DNA samples of simulated Type 2 diabetes and actual samples of Type 2 diabetes.

Description

technical field [0001] The invention relates to a DNA microarray chip detection method for single nucleotide polymorphism related to type II diabetes, belonging to the technical field of DNA microarray chips. Background technique [0002] Type Ⅱ diabetes is a polygenic heterogeneous disease caused by genetic and environmental factors. Studying its pathogenesis from the perspective of genetics is very important for the screening, diagnosis and intervention of prediabetes (A genome-wide association study identified novel risk loci for type 2diabetes. Nature. 2007, 445, 881-885; Genome-Wide Association Analysis Identifies Loci for Type 2Diabetes and Triglyceride Levels. Science, 2007, 316, 1331-1336.). In the study of the genetic mechanism of complex diseases, single nucleotide polymorphisms (single nucleotide polymorphisms, SNPs) analysis can provide a convenient and effective way for the study of the genetic mechanism of complex traits. Screening oligonucleotide probes that...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6837C12Q1/6883C12Q2600/156C12Q2563/137
Inventor 王振新李桃邢雪峰邬东阳
Owner CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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