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Method for simultaneously detecting 32 single nucleotide polymorphism (SNP) locus genotypes

A single nucleotide polymorphism, gene technology, applied in the field of medical molecular biology, can solve the problems of poor sensitivity, easy missed diagnosis, misdiagnosis, difficult to popularize and so on

Inactive Publication Date: 2010-10-06
GENERAL HOSPITAL OF PLA
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Electrocardiogram is a common method for the diagnosis and treatment of cardiovascular diseases. It has the advantages of simplicity, rapidity, low price, non-invasiveness and good repeatability, but its sensitivity is poor, the false positive rate is high, and it is easy to miss and misdiagnose.
Two-dimensional echocardiography, radionuclide myocardial perfusion imaging, and electron beam CT have high sensitivity and specificity for the diagnosis of coronary heart disease, but the equipment is expensive. Although coronary angiography is the gold standard for the diagnosis of coronary heart disease Standards are difficult to popularize because of their technical complexity and certain invasiveness
These methods can only detect one or a few single nucleotide polymorphisms, and are not suitable for the detection of multiple single nucleotide polymorphisms, multiple types of single nucleotide polymorphisms and large batches of samples

Method used

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  • Method for simultaneously detecting 32 single nucleotide polymorphism (SNP) locus genotypes
  • Method for simultaneously detecting 32 single nucleotide polymorphism (SNP) locus genotypes
  • Method for simultaneously detecting 32 single nucleotide polymorphism (SNP) locus genotypes

Examples

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Embodiment 1

[0069] Example 1 The genotypes of 32 single nucleotide polymorphism sites in the Chinese Han population were simultaneously detected by the method of the present invention.

[0070] (1) Designing 32 pairs of primers and 32 extended primers for sequencing and hybridization for amplifying fragments containing the target single nucleotide polymorphism site, see sequence 1 to sequence 96 for details. .

[0071] (2) Dilute and mix the PCR primers so that the final concentration is 2.5 μM, construct the PCR primer pool and amplify the target fragment. The PCR reaction system for every 96 people is:

[0072]

[0073] Reagent Components Volume (μL)

[0074]

[0075] Primer Pool (2.5μM) 11.2

[0076] Deoxyribonucleotides (2.5mM) 20

[0077] 10x PCR Buffer II 56.2

[0078] Magnesium chloride (25mM) 112.6

[0079] Gold Amplification Enzyme (5U / μL) 11.2

[0080] Double distilled water 126

[0081] ...

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Abstract

The invention relates to a method for simultaneously detecting 32 labeled single nucleotide polymorphism (SNP) loci. In the method, adducin-1, platelet endothelial cell adhesion molecule-1 (PECAM-1), C-reactive protein (CRP), endothelial constitutive nitric oxide synthase (ecNOS), plasma cell membrane glycoprotein-1 (PC-1), L-selectin, G-protein beta3 subunit gene, angiotensin I converting enzyme (ACE), angiotensin II type 1 receptor gene, proangiotensin, methylenetetrahydrofolate reductase (MTHFR) and hepatic lipase gene which are published on an international haplotype diagram engineering database are selected for detecting 32 labeled single nucleotide polymorphisms of the Han nationality in China; three primers are designed for each single nucleotide polymorphism (two PCR amplification primers and one extension primer); and the single nucleotide polymorphism loci are subjected to genotyping through polymerase chain reaction (PCR) amplification, extension and hybridization and by applying an SNP stream genotyping system. The detection method has the advantages of quickness, accuracy, high throughput, simple operation, microscale and the like.

Description

technical field [0001] The invention relates to a method for simultaneously detecting 32 single nucleotide polymorphism sites. It belongs to the field of medical molecular biology. Background technique [0002] In recent years, with the improvement of people's living standards and changes in lifestyles, coronary heart disease has become a major disease that endangers people's health and lives in our country. . Its pathogenesis is the result of the joint action of genetic factors and environmental factors. Coronary heart disease is mainly caused by atherosclerotic plaques. The formation of atherosclerosis is affected by many factors. A large number of clinical experiments have confirmed that blood lipid levels play an important role in the occurrence and development of atherosclerosis. Dyslipidemia is a major risk factor for coronary heart disease and atherosclerosis. In addition, the inflammatory response runs through the entire process of atherosclerosis, and the acti...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 田亚平张阳东
Owner GENERAL HOSPITAL OF PLA
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