Detection method and kit for BUD13-ZNF259 gene mononucleotide polymorphism and application thereof

1. The technology of BUD13-ZNF259 and single nucleotide polymorphism is applied in the direction of recombinant DNA technology, measurement/inspection of microorganisms, biochemical equipment and methods, etc., which can solve the problems of non-disclosure and achieve high accuracy and method Simple and easy to operate effect

Inactive Publication Date: 2015-06-03
NINGBO UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, at present, there are no relevant research reports on the kit used to detect the correlation between BUD13-ZNF259 gene rs964184 single nucleotide polymorphism (SNP) and coronary heart disease at home and abroad

Method used

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  • Detection method and kit for BUD13-ZNF259 gene mononucleotide polymorphism and application thereof
  • Detection method and kit for BUD13-ZNF259 gene mononucleotide polymorphism and application thereof
  • Detection method and kit for BUD13-ZNF259 gene mononucleotide polymorphism and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Collection of research objects: A total of 874 patients with coronary heart disease were collected from Ningbo Lihuili Hospital, Ningbo Yinzhou People's Hospital, and Zhejiang Hangzhou Second Hospital, including 625 males and 249 females, with an average age of 62 years old; Among these inpatients, patients with less than 50% stenosis of each coronary artery were collected as the control group, a total of 776 cases, including 424 males and 352 females, with an average age of 58 years old. None of the participants had myocardial infarction, congenital heart disease, liver or kidney disease.

[0042] 1. Extraction of Serum DNA

[0043] Use Lab-Aid 820 automatic nucleic acid extractor (Xiamen Zhishan Biotechnology Co., Ltd., China, 500ul system) to extract whole blood genomic DNA from the sample, and then detect the concentration of the obtained DNA with a nucleic acid protein analyzer.

[0044] 2. Primer design: According to the sequence near the target SNP rs964184 down...

Embodiment 2

[0058] 1. Genotyping results

[0059] The invention judges the genotype according to the Tm value, LL homozygote, that is, GG: 76±0.5°C, HH homozygote, that is, CC: 80±0.5°C; LH heterozygote, that is, CG, Tm is 76±0.5°C and 80±0.5°C .

[0060] The genotype was detected by the Tm-shift method ( Figure 1-3 ), among 1650 patients, GG accounted for 5.33% (88 / 1650); CG accounted for 35.94% (593 / 1650); CC accounted for 58.72% (969 / 1650).

[0061] 2. Correlation between rs964184 and coronary heart disease (P<0.05 means there is a correlation)

[0062] By comparing patients and controls, we found that: as shown in Table 1, the risk gene G gene of rs964184 in the case group was significantly higher than that in the control group, and its inheritance mode was dominant inheritance.

[0063] Table 1 Results of genotype and allele distribution of rs964184

[0064]

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Abstract

The invention relates to a method for detecting BUD13-ZNF259 gene rs964184 mononucleotide polymorphism by a Tm-shift process. The method comprises the following steps: respectively adding GS-rich sequence to the 5' end of a specific primer, and pairing base of the 3' terminal with allele mutant of SNP (single-nucleotide polymorphism); and in the presence of the primer, carrying out PCR (polymerase chain reaction) amplification on DNA (deoxyribonucleic acid) extracted from the serum sample in a fluorescence PCR instrument, analyzing the PCR product by using a melting curve, and judging the genotype according to the Tm value of the PCR product. The primer can be used for preparing a kit for detecting susceptibility of coronary heart disease. The Tm-shift process is simple to operate, has the advantages of high accuracy, favorable repetitiveness and low cost, and can be used for promoting early prevention of diseases and providing references for drug therapy.

Description

technical field [0001] The invention relates to a detection kit for auxiliary diagnosis of coronary heart disease, in particular to a detection kit for detecting the correlation between BUD13-ZNF259 gene rs964184 single nucleotide polymorphism (SNP) and coronary heart disease and its application. Background technique [0002] Coronary heart disease is also called ischemic heart disease (coronary heart disease, CHD), which involves atherosclerosis in the arteries supplying myocardial blood, that is, coronary atherosclerosis leads to stenosis of the blood vessel lumen or plaque formation or even rupture, complete blockage, limiting Or completely interrupt the blood supply of the myocardium, causing clinically a series of severe myocardial ischemic diseases such as angina pectoris and myocardial infarction. Coronary heart disease is one of the most serious diseases threatening human health today, and it is the main killer of human health. With the rapid development of the econ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6883C12Q1/6858C12Q2600/156
Inventor 李奕润段世伟叶华丹周安楠
Owner NINGBO UNIV
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