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Coronary disease testing method and reagent kit

A kit and in vitro detection technology, applied in the fields of molecular biology and medicine, can solve the problem that the correlation between PON3 gene SNP and atherosclerosis and/or coronary heart disease is not confirmed, and the relationship between PON3 gene and atherosclerotic coronary heart disease is not confirmed Relevance, etc.

Inactive Publication Date: 2008-10-29
CHINESE NAT HUMAN GENOME CENT AT SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0020] Although there have been many studies on the relationship between various gene polymorphisms and atherosclerosis and / or coronary heart disease, there is no report on the correlation between PON3 gene and atherosclerosis and / or coronary heart disease, let alone the correlation of PON3 gene. A report on the correlation between SNP and atherosclerosis and / or coronary heart disease

Method used

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  • Coronary disease testing method and reagent kit
  • Coronary disease testing method and reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] 1.1 Research object

[0069] Among the patients diagnosed with atherosclerosis by coronary angiography, the patients with 2 or more lesions and a stenosis of more than 50% were selected as cases. In addition, normal people without atherosclerosis diagnosed by coronary angiography were selected as controls, and the age, sex, and place of origin were matched with the case group as much as possible.

[0070] Peripheral blood samples were randomly collected from 190 patients and 191 normal control individuals on the basis of informed consent.

[0071] 1.2 Experimental methods and results

[0072] 1.2.1 DNA extraction

[0073] DNA was extracted from human peripheral blood samples by the conventional phenol-chloroform method, and the concentration was corrected to 20ng / ul for conventional PCR amplification.

[0074] 1.2.2 Design of PCR and sequencing primers

[0075] According to the genome sequence of PON3 in GenBank, the following primers were designed and synthesized. ...

Embodiment 2

[0089] Atherosclerosis and / or Coronary Heart Disease Susceptibility Detection Kit

[0090] As described in Example 1, the T→C mutation at position 301 in SEQ ID NO: 1 is closely related to atherosclerosis and / or coronary heart disease. Therefore, based on this mutation, PON3 gene-specific primers can be designed to amplify and detect with the patient's DNA as a template.

[0091] Prepare a test kit (100 person-times), which contains:

[0092]

[0093] 100 people were randomly selected to form a test group, including subjects who were unknown whether they had atherosclerosis, patients who were known to suffer from atherosclerosis and / or coronary heart disease, and normal people who had been tested without atherosclerosis.

[0094] Take 3ml of peripheral blood from the subject to be tested in the test group, and use conventional methods (or use a specific kit) to extract DNA from the blood. Dilute the PCR primers in the atherosclerosis and / or coronary heart disease detectio...

Embodiment 3

[0100] Auxiliary testing for susceptibility to atherosclerosis and / or coronary heart disease

[0101] The test in Example 2 was repeated, with the difference that 80 people (whose symptoms of atherosclerosis were not known before the test) were randomly selected for testing.

[0102] Prepare a test kit (100 person-times), which contains:

[0103]

[0104] Take 3ml of peripheral blood from the subject to be tested, and use conventional methods (or use a specific kit) to extract DNA from the blood. Dilute the PCR primers in the atherosclerosis and / or coronary heart disease detection kit to 1μmol / μl and use the extracted DNA as a template to perform PCR reaction with the provided primers. After purification of PCR products, ABI-PRISM TM The 377 DNA sequencer performs bidirectional sequencing by the fluorescent labeling end termination method, and the sequence interpretation and SNP confirmation are performed with Polyphred software.

[0105] The results also confirmed that...

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Abstract

The invention discloses a method for detecting atherosclerosis and / or coronary heart disease susceptibility, which includes detecting whether variation exists in individual paraoxonase 3 (PON3), transcript and / or albumen in comparison with normal. The existence of variation shows that the possibility that an individual contracts atherosclerosis and / or coronary heart disease is higher than that of a normal person. The invention also discloses a corresponding detection kit.

Description

technical field [0001] The present invention relates to the fields of molecular biology and medicine. More specifically, it relates to the single nucleotide polymorphism (single nucleotide polymorphism, SNP) of paraoxonase 3 (Paraoxonase3, PON3) and its correlation with atherosclerosis and / or coronary heart disease. The present invention also relates to methods and kits for detecting these SNPs. Background technique [0002] Atherosclerosis is a common progressive arterial disease. The lesion mainly involves medium-sized muscular arteries. Lipid deposition in the arterial intima and proliferation of smooth muscle cells form localized plaques, which can harden the arterial wall. Plaque rupture leads to thrombus, embolism, hemorrhage, partial or complete occlusion of the involved lumen, and the clinical manifestation is the occurrence of atherosclerotic vascular complications. The most common and serious vascular complications in elderly patients are atherosclerosis and / or O...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 黄薇王颖王海丰王一金力
Owner CHINESE NAT HUMAN GENOME CENT AT SHANGHAI
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