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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 unconfirmed correlation between PON3 gene and atherosclerotic coronary heart disease, unconfirmed PON3 gene SNP and atherosclerosis and/or coronary heart disease issues of relevance

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 patients diagnosed with atherosclerosis by coronary angiography, select patients with 2 or more lesions and more than 50% lesion stenosis as the case. In addition, normal people who were diagnosed as having no atherosclerosis by coronary angiography were taken as controls, and the age, gender and hometown were selected to match the case group as much as possible.

[0070] On the basis of informed consent, peripheral blood samples of 191 patients and 191 normal control individuals were randomly collected.

[0071] 1.2 Experimental methods and results

[0072] 1.2.1 DNA extraction

[0073] The DNA was extracted from human peripheral blood samples by the conventional phenol-chloroform method, and the concentration was adjusted to 20ng / ul, and then used 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 syn...

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, specific primers for the PON3 gene can be designed for amplification using the patient's DNA as a template for detection.

[0091] Prepare a kit (100 visits), which contains:

[0092]

[0093] A test group consisting of 100 people was randomly selected, including subjects who are not known to have atherosclerosis, patients who are known to have atherosclerosis and / or coronary heart disease, and normal people who have not been tested for atherosclerosis.

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

Embodiment 3

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

[0101] The test of Example 2 was repeated, the difference is that 90 people were randomly selected (we don't know whether they have atherosclerosis symptoms before the test) for testing.

[0102] Prepare a kit (100 visits), which contains:

[0103]

[0104] 3ml of peripheral blood of the subject to be tested is drawn, and DNA is extracted from the blood using conventional methods (or using a specific kit). 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 a PCR reaction with the provided primers. After the PCR product is purified, use ABI-PRISM TM 377 DNA sequencer performs two-way sequencing with fluorescent labeling terminal termination method, and uses Polyphred software for sequence interpretation and SNP confirmation.

[0105] The results also confirmed that the proportion of athero...

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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 invention relates to the fields of molecular biology and medicine. More specifically, it relates to single nucleotide polymorphism (SNP) of paraoxonase 3 (Paraoxonase 3, 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 disease mainly involves medium-sized muscular arteries, lipid deposition in the arterial intima, proliferation of smooth muscle cells, and the formation of localized plaques that can harden the arterial wall. Plaque rupture leads to thrombosis, embolism, hemorrhage, and partial or complete occlusion of the involved lumen. The clinical manifestation is the occurrence of atherosclerotic vascular complications. The most common and serious vascular complications in elderly patients are atherosclerosis and / Or coronary heart disease (unstable an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
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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