CN101294200A - Coronary heart disease detection method and kit - Google Patents

Abstract

The present invention discloses a method for detecting susceptibility to atherosclerosis and/or coronary heart disease, which comprises detecting the presence or absence of platelet-derived growth factor receptor alpha peptide (PDGFRA), transcript and/or protein in an individual compared with normal The existence of variation indicates that the individual is more likely to suffer from atherosclerosis and/or coronary heart disease than the normal population. The invention also discloses a corresponding detection kit.

Description

Coronary heart disease detection method and kit

technical field

The present invention relates to the fields of molecular biology and medicine. More specifically related to platelet-derived growth factor receptor alpha peptide (platelet-derived growth factor receptor, alpha polypeptide, PDGFRA) single nucleotide polymorphism (single nucleotide polymorphism, SNP) and its association with atherosclerosis and/or Correlation with coronary heart disease. The present invention also relates to methods and kits for detecting these SNPs.

Background technique

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 Or coronary heart disease (unstable angina and myocardial infarction) and stroke.

The occurrence and development of atherosclerotic lesions mainly undergo four progressive pathological processes:

1. Intimal changes in the early stage of lipid infiltration: endothelial injury

2. Lipid streaks: accumulation of foam cells and T lymphocytes, hyperplasia of smooth muscle cells.

3. Fibrous plaque: smooth muscle fibrosis, forming a fibrous cap.

4. Compound lesions: plaque calcification, rupture, ulceration, and medial lesions.

The etiology and pathogenesis of atherosclerosis are very complex. It is a series of interactions among arterial wall cells, extracellular matrix, blood components (especially monocytes, platelets and LDL), local hemodynamics, environment and genetic factors. The result of complex effects, so the etiology is not a single factor.

As a polygenic disease, atherosclerosis not only involves multiple genes with different roles, but also involves gene-gene and gene-environment interactions. Now there are many theories about the pathogenesis of atherosclerosis, such as the theory of infiltration of lipids, the theory of vascular intima injury, the theory of inflammatory response, the theory of oxidation-antioxidation and the theory of thrombosis. All these theories involve different biological pathways, but none of them can be fully explained in genetics and molecular biology.

Over the years, based on the long-term research on the risk factors of atherosclerosis, scientists have carried out a lot of work on these risk factors. So far, certain insertions, deletions and mutations of angiotensin I converting enzyme (ACE) and MTHFR have been found to be potential risk factors for atherosclerosis and/or coronary heart disease. The former is one of the enzymes that play an important role in blood pressure regulation, and the latter is an enzyme related to cysteine metabolism.

In addition, German scientists have recently confirmed that the expression level of AT1 receptors is closely related to the process of vascular damage: ACE inhibitors will reduce the activation of AT1 receptors, thereby improving poor vascular function and effectively inhibiting the occurrence and development of atherosclerosis.

However, for a long time, domestic and foreign studies have focused more on the mutation and expression of related genes in lipid metabolism pathways, such as: paraoxygenase, lipoprotein lipase, liver lipase, etc. In the study of endogenous nitric oxide synthase (eNOS), some research work has shown that eNOS is closely related to atherosclerosis. Another American scientist proposed the role of inflammatory mechanism in the formation of atherosclerosis, which was confirmed in mouse experiments.

In recent years, the concept of single nucleotide polymorphism (SNP) and haplotype (haplotype) has been widely used in the study of polygenic diseases, which has opened up a new era for the study of the mechanism of the occurrence and development of atherosclerosis at the molecular level. train of thought. At the same time, the continuous emergence of high-throughput and low-cost SNP detection methods also provides the possibility for large-scale and rapid SNP typing.

Japanese scientists used a large-scale case-control study to conduct association analysis on more than 90,000 related gene SNPs, and found a 50KB haplotype region that is closely related to myocardial infarction, including LTA (lymphotoxin-α), NFKBIL1 and BAT1 and so on for multiple SNP sites.

In another study, although S290N and N562D in P-selectin (P-selectin) were not independently associated with myocardial infarction, their haplotypes were significantly associated with myocardial infarction.

In addition, there are racial and geographical differences in the presence or absence of SNPs and allele frequencies, which suggests the existence of genetic heterogeneity in polygenic diseases, that is, the same disease or trait may be caused by different genetic factors in different populations of. In the study of the blood coagulation factor V gene, the VLeiden mutation reported abroad that was associated with the incidence of myocardial infarction in white women was not found in the Chinese population, but a new SNP-G/A1628 (Arg485Lys) mutation was found that was associated with APCR and Atherosclerosis and/or coronary heart disease are related, and the coagulation factor V gene may be a susceptibility gene for atherosclerosis in the Chinese population.

In addition, the differences in the types and frequencies of SNPs and their haplotypes in different populations may be closely related to the different responses of different populations to drugs and environmental factors.

In the past 50 years, large-scale epidemiological surveys and genetic studies have done a series of studies on coronary atherosclerosis, a complex disease, and found that some susceptibility genes are related to atherosclerosis, such as ABCA1 , CYBA, ApoE, LDLR, etc. More atherosclerosis-related ones continue to be discovered and validated.

Atherosclerosis and/or coronary heart disease is a polygenic disease caused by the interaction of genetic factors and environmental factors. It has become a current study to find the related genes and then clarify the genetic mechanism of atherosclerosis and/or coronary heart disease. hotspots.

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 PDGFRA gene and atherosclerosis and/or coronary heart disease, let alone the correlation of PDGFRA gene. Reports of association of SNPs with atherosclerosis and/or coronary heart disease.

In summary, in order to diagnose atherosclerosis and/or coronary heart disease as early as possible, there is an urgent need in the field to find susceptibility genes for atherosclerosis and/or coronary heart disease, and to develop methods for detecting atherosclerosis and/or coronary heart disease and kits.

Contents of the invention

The object of the present invention is to provide a method and detection kit for auxiliary diagnosis (especially early diagnosis) of atherosclerosis and/or coronary heart disease.

Another object of the present invention is to provide a new method for treating atherosclerosis and/or coronary heart disease.

In a first aspect of the present invention, a method of diagnosing susceptibility to atherosclerosis and/or coronary heart disease in an individual is provided, comprising the steps of:

detecting the individual's PDGFRA gene, transcript and/or protein, and comparing it with normal PDGFRA gene, transcript and/or protein,

A difference indicates that the individual is more likely to suffer from atherosclerosis and/or coronary heart disease than the normal population.

In another preferred example, the method detects the gene or transcript of PDGFRA, and compares the difference with the normal PDGFRA nucleotide sequence.

In another preferred example, the difference is the following single nucleotide polymorphism:

301 bits A→G;

Wherein, the nucleotide position numbers are based on SEQ ID NO:1.

In a second aspect of the present invention, there is provided a method for detecting whether a single nucleotide polymorphism of platelet-derived growth factor receptor alpha peptide (PDGFRA) exists in a sample, comprising the steps of:

(a) amplifying the PDGFRA gene of the sample with the PDGFRA gene-specific primers to obtain an amplified product; and

(b) Detect whether the following single nucleotide polymorphisms exist in the amplification product:

301 bits A→G;

Wherein, the nucleotide position numbers are based on SEQ ID NO:1.

In another preferred example, the gene-specific primers have the sequences of SEQ ID NO: 2 and 3.

In another preferred example, the length of the amplified product is 100-2000bp, and contains the 301st position in SEQ ID NO:1.

In a third aspect of the present invention, a kit for detecting atherosclerosis and/or coronary heart disease is provided, which includes primers for specifically amplifying the PDGFRA gene or transcripts, more preferably, said primers amplify The length is 100-2000bp, and contains the 301st amplification product in SEQ ID NO:1.

In another preferred embodiment, the kit also contains reagents selected from the following group:

(a) a probe that binds to the mutation at position 301 in SEQ ID NO: 1;

(b) Recognizing the mutant restriction enzyme at position 301 in SEQ ID NO:1.

In another preferred example, the mutation is selected from the following single nucleotide polymorphisms:

301 bits A→G;

Wherein, the nucleotide position numbers are based on SEQ ID NO:1.

Detailed ways

After intensive and extensive research, the inventors have determined and analyzed the SNPs of a large number of candidate genes. For the first time, it was discovered and proved that the genome sequence of PDGFRA is closely related to atherosclerosis and/or coronary heart disease. The results of the association study showed that the SNP at position 301 in SEQ ID NO: 1 of PDGFRA (position 301 A→G)( Denoted as RS36104023) there is a significant difference (P<0.05) in the distribution of cases and controls, so it can be used as a specific SNP for auxiliary detection of atherosclerosis and/or coronary heart disease (or its susceptibility). The present invention has been accomplished on this basis.

PDGFRA gene

The sequence of platelet-derived growth factor receptor α peptide (platelet-derived growth factor receptor, alpha polypeptide, PDGFRA) is known, its detailed sequence and some related information can be found at the website http://www.ncb i.nlm.nih. gov/Genebank/; http://www.ncbi.nlm.nih.gov/SNP. For convenience, the nucleotide sequence relevant to the SNP of the present invention in PDGFRA is given in SEQ ID NO: 1.

The PDGFRA gene is a receptor for members of the platelet-derived growth factor (PDGF) family. It is a tyrosine kinase that binds to corresponding growth factors and transmits signals that promote cell growth. High expression of PDGFRA gene can cause hyperplasia of eosinophils, thus triggering a series of downstream adverse reactions.

The inventors sequenced almost the entire region of the PDGFRA gene and found many SNPs, most of which were not associated with susceptibility to atherosclerosis and/or coronary heart disease. However, association studies showed that in SEQ ID NO: 1 The 301st position A→G is a SNP highly associated with the susceptibility of atherosclerosis and/or coronary heart disease.

Specifically, the present invention discloses the single nucleotide polymorphism (SNP) of No. 15 intron of PDGFRA gene and the correlation between the polymorphism and coronary atherosclerosis. The SNP of the present invention is the A/G polymorphism of the 301st position of the sequence shown in SEQ ID NO: 1, which is located in the No. 15 intron of the human PDGFRA gene, and the heterozygous AG in the coronary atherosclerosis patient population The frequency is higher than that in the normal control population.

Based on the new discovery of the present invention, the PDGFRA protein or polypeptide has many new uses. These uses include (but are not limited to): for auxiliary diagnosis of atherosclerosis and/or coronary heart disease, or for screening substances that promote the function of PDGFRA protein, such as antibodies, polypeptides or other ligands.

On the other hand, the present invention also includes polyclonal antibodies and monoclonal antibodies specific to human PDGFRA DNA or polypeptides encoded by its fragments, especially monoclonal antibodies. Here, "specificity" means that the antibody can bind to human PDGFRA gene product or fragment. Preferably, it refers to those antibodies that can bind to human PDGFRA gene products or fragments but do not recognize and bind to other irrelevant antigen molecules. Antibodies in the present invention include those molecules that can bind to and inhibit human PDGFRA protein, as well as those that do not affect the function of human PDGFRA protein.

The present invention includes not only complete monoclonal or polyclonal antibodies, but also immunologically active antibody fragments, such as Fab' or (Fab) ₂ fragments; antibody heavy chains; antibody light chains; genetically engineered single-chain Fv molecules; or chimeric antibodies.

Antibodies of the present invention can be prepared by various techniques known to those skilled in the art. For example, a purified human PDGFRA gene product, or an antigenic fragment thereof, can be administered to an animal to induce polyclonal antibody production. Similarly, cells expressing human PDGFRA protein or antigenic fragments thereof can be used to immunize animals to produce antibodies. Various adjuvants can be used to enhance the immune response, including but not limited to Freund's adjuvant and the like.

Antibodies of the invention may also be monoclonal antibodies. Such monoclonal antibodies can be prepared using hybridoma technology. The antibodies of the present invention include antibodies capable of blocking the function of human PDGFRA protein and antibodies that do not affect the function of human PDGFRA protein. Various types of antibodies of the present invention can be obtained by conventional immunization techniques using fragments or functional regions of human PDGFRA gene products. These fragments or functional regions can be prepared using recombinant methods or synthesized using a polypeptide synthesizer. Antibodies that bind to unmodified forms of the human PDGFRA gene product can be produced by immunizing animals with gene products produced in prokaryotic cells (e.g., E. coli); antibodies that bind to post-translationally modified forms (such as glycosylated or phosphorylated Proteins or polypeptides), which can be obtained by immunizing animals with gene products produced in eukaryotic cells (such as yeast or insect cells).

The antibody against human PDGFRA protein can be used in immunohistochemical techniques to detect the amount and/or mutation of human PDGFRA protein in biopsy specimens. A preferred anti-PDGFRA antibody is an antibody that does not recognize normal PDGFRA but recognizes mutant PDGFRA, or an antibody that recognizes normal PDGFRA but not mutant PDGFRA. Using these antibodies, the detection of atherosclerosis and/or coronary heart disease susceptibility at the protein level can be conveniently performed.

Using the PDGFRA protein of the present invention, substances that interact with the PDGFRA protein, such as inhibitors, agonists or antagonists, can be screened out through various conventional screening methods.

The invention also relates to a diagnostic test method for quantitatively and locally detecting human PDGFRA protein level. These assays are well known in the art and include ELISA and the like.

A method for detecting whether there is PDGFRA protein in a sample is to use a specific antibody for PDGFRA protein to detect, which includes: contacting the sample with a specific antibody for PDGFRA protein; observing whether an antibody complex is formed, which means that the antibody complex is formed PDGFRA protein is present in the sample.

The polynucleotide of PDGFRA protein can be used for auxiliary diagnosis of diseases related to PDGFRA protein. In terms of diagnosis, the polynucleotide of PDGFRA protein can be used to detect the expression of PDGFRA protein or the abnormal expression of PDGFRA protein in a disease state. For example, the PDGFRA DNA sequence can be used for hybridization of biopsy specimens to determine the abnormal expression of PDGFRA protein. Hybridization techniques include Southern blotting, Northern blotting, in situ hybridization, and the like. These technical methods are all open and mature technologies, and relevant kits are available from commercial sources. Part or all of the polynucleotides of the present invention can be immobilized as probes on microarrays or DNA chips (also known as "gene chips") for analysis of differential expression of genes in tissues and gene diagnosis. RNA-polymerase chain reaction (RT-PCR) in vitro amplification with PDGFRA protein-specific primers can also detect the transcripts of PDGFRA protein.

Detection can be against cDNA or genomic DNA. The form of PDGFRA protein mutation includes point mutation, translocation, deletion, recombination and any other abnormality compared with the normal wild-type PDGFRA DNA sequence. Mutations can be detected using established techniques such as Southern blotting, DNA sequence analysis, PCR and in situ hybridization. In addition, mutations may affect protein expression, so Northern blotting and Western blotting can be used to indirectly determine whether a gene has a mutation.

The most convenient method for detecting the SNP of the present invention is to amplify the PDGFRA gene of the sample with PDGFRA gene-specific primers to obtain the amplified product; then detect whether there is the following single nucleotide polymorphism in the amplified product: 301 position A → G, wherein the nucleotide position numbering is based on SEQ ID NO:1.

It should be understood that after the present invention first revealed the correlation between the SNP of the PDGFRA gene and atherosclerosis and/or coronary heart disease, those skilled in the art can easily design an amplification method that can specifically amplify the position containing the SNP The product, and then determine whether there is 301 A→G by sequencing and other methods. Usually, the length of the primer is 15-50bp, preferably 20-30bp. Although it is preferred that the primer is completely complementary to the template sequence, those skilled in the art know that it can also be specifically amplified (that is, only amplify the desired fragment). Kits containing these primers and methods using these primers are within the scope of the present invention, as long as the amplified product amplified by the primers contains the corresponding position of the SNP of the present invention. A preferred primer pair has the sequences of SEQ ID NO: 2 and 3.

Although the length of the amplified product is not particularly limited, generally the length of the amplified product is 100-2000 bp, preferably 150-1500 bp, more preferably 200-1000 bp. These amplified products should all contain the 301st position in SEQ ID NO:1.

Because the SNP of the present invention has a very high correlation with atherosclerosis and/or coronary heart disease, it can not only be used for early auxiliary diagnosis of atherosclerosis and/or coronary heart disease, but also can prevent some carriers in the future Take reasonable preventive measures (such as taking corresponding control in diet) before the onset of the disease, so as to improve the survival period and quality of life of the carrier, so it has extremely important application value and social benefits. Of course, the final diagnosis should be confirmed by routine testing methods.

Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental method that does not indicate specific conditions in the following examples, usually according to conventional conditions such as Sambrook et al., molecular cloning: the conditions described in the laboratory manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the manufacturer's instructions suggested conditions.

Example 1

1.1 Research object

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.

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

1.2 Experimental methods and results

1.2.1 DNA extraction

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.

1.2.2 Design of PCR and sequencing primers

According to the genome sequence of PDGFRA in GenBank, the following primers were designed and synthesized. The specific primers are shown in Table 1 below.

Table 1 Primer sequence list

Primer name Sequence (5'-3') SEQ ID NO: sense primer gtgatccact tagacctata aaatg 2 antisense primer tgacaaagat gggaaagact agcgg 3

1.2.3 PCR amplification of PDGFRA gene

Using the extracted DNA as a template, PCR amplification was performed on a GeneAmp 9700 PCR instrument with the Touchdown program using Taq enzyme. The reaction conditions are: pre-denaturation at 94°C for 2 minutes, denaturation at 94°C for 30 seconds, annealing at 63°C for 40 seconds, extension at 72°C for 40 seconds, a total of 10 cycles, and the annealing temperature decreases by 0.5°C for each cycle; after denaturation at 94°C for 30 seconds, Anneal at 58°C for 40 seconds, extend at 72°C for 40 seconds, a total of 30 cycles; finally extend at 72°C for 7 minutes. PCR amplification products were verified by agarose gel electrophoresis. As a result, an amplification product of PDGFRA was obtained.

1.2.4 SNP discovery and detection

After the PCR products were purified by Resin resin, the ABI-PRISM ^TM 377 DNA sequencer (appliedbiosystems (ABI)) was used for bidirectional sequencing with fluorescent labeling end termination method, and Polyphred software (University of Washington, USA http://droog. mbt.washington.edu/Polyphred.html) for sequence interpretation and SNP confirmation.

As a result, several SNPs were found to exist, including the following SNP: A→G at position 301 in SEQ ID NO:1.

1.2.5 SNP genotyping and association analysis

SNP genotyping by direct one-way sequencing. That is, typing and association analysis were performed in patients with atherosclerosis and/or coronary heart disease and normal controls.

Descriptive statistical analysis was performed on the genotyping results, and the chi-square test was performed on the contingency table. See if genotypes differ between patients and controls.

The analysis results showed that in a total of 190 patients and 191 normal controls, there was a significant difference in the 301 SNP of PDGFRA (denoted as RS36104023) between the patient group and the control group: the single intron 15 of the PDGFRA gene Among the nucleotide polymorphism sites, the frequency of heterozygous AG was 37.4% in the patient population and 21.5% in the control group, and there was a significant difference between the two (p=0.001).

The above results confirmed that the SNP at position 301 in SEQ ID NO: 1 is correlated with the occurrence of atherosclerosis and/or coronary heart disease.

Example 2

Atherosclerosis and/or Coronary Heart Disease Susceptibility Detection Kit

As described in Example 1, the mutation of A→G at position 301 in SEQ ID NO: 1 is closely related to atherosclerosis and/or coronary heart disease. Therefore, PDGFRA gene-specific primers can be designed based on this mutation and detected by amplification using the patient's DNA as a template.

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

A test group composed of 100 people was randomly selected, including subjects who were unknown whether they had atherosclerosis, patients who were known to have atherosclerosis and/or coronary heart disease, and normal people who had been tested without atherosclerosis.

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 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 products were purified, ABI-PRISM ^TM 377 DNA sequencer was used for bidirectional sequencing by fluorescence-labeled end termination method, and Polyphred software was used for sequence interpretation and SNP confirmation.

Alternatively, the chromatographic analysis of the amplified product and the normal control is performed with a denaturing high-performance liquid chromatograph (DHPLC), and A→G at position 301 can also be detected.

Test results:

For subjects with 301 positions A→G in PDGFRA, it is further tested by conventional methods to confirm whether they have atherosclerosis. The test results show that the susceptibility ratio of atherosclerosis and/or coronary heart disease of the test subjects (especially AG heterozygotes) containing 301 A→G is significantly higher than that of the normal population (at least 1 times higher than that of AA at this site). ).

Therefore, this indicates that auxiliary detection and/or early diagnosis of atherosclerosis can be performed by detecting A→G at position 301 of PDGFRA.

Example 3

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

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.

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

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 a PCR reaction with the provided primers. After the PCR products were purified, ABI-PRISM ^TM 377 DNA sequencer was used for bidirectional sequencing by fluorescence-labeled end termination method, and Polyphred software was used for sequence interpretation and SNP confirmation.

The results also confirmed that the proportion of atherosclerosis in the test subjects with 301 A→G (especially the AG heterozygote) was significantly higher than that in the test subjects with AA at this site.

All documents mentioned in this application are incorporated by reference in this application as if each were individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

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Claims (10)

1. whether a vitro detection sample exists the method for the single nucleotide polymorphism of platelet derived growth factor receptor α peptide (PDGFRA), it is characterized in that, comprises step:

(a) with the PDGFRA gene of PDGFRA gene-specific primer amplification sample, obtain amplified production; With

(b) detect whether there is following single nucleotide polymorphism in the amplified production:

301 A → G;

Wherein, the nucleotide position numbering is based on SEQ ID NO:1.

2. the method for claim 1 is characterized in that, described gene-specific primer has the sequence of SEQID NO:2 and 3.

3. the method for claim 1 is characterized in that, the length of described amplified production is 100-2000bp and contains among the SEQ ID NO:1 the 301st.

4. test kit that detects atherosclerosis and/or coronary heart disease, it is characterized in that, it comprises the primer of specific amplification PDGFRA gene or transcript, and described primer amplification goes out length to be 100-2000bp and to contain among the SEQ ID NO:1 the 301st amplified production.

5. test kit as claimed in claim 4 is characterized in that, it also contains the reagent that is selected from down group:

(a) with SEQ ID NO:1 in the 301st sudden change bonded probe;

(b) the 301st sudden change restriction enzyme among the identification SEQ ID NO:1.

6. test kit as claimed in claim 5 is characterized in that, described sudden change is following single nucleotide polymorphism:

301 A → G;

Wherein, the nucleotide position numbering is based on SEQ ID NO:1.

7. test kit as claimed in claim 4 is characterized in that described primer has the sequence of SEQ ID NO:2 and 3.

8. method that the atherosclerosis and/or the coronary disease susceptibility of individuality are diagnosed is characterized in that it comprises step:

Detect this individual PDGFRA gene, transcript and/or albumen, and compare with normal PDGFRA gene, transcript and/or albumen,

Wherein, the possibility that there are differences with regard to showing this individuality trouble atherosclerosis and/or coronary heart disease is higher than normal population.

9. method as claimed in claim 8 is characterized in that, detection be gene or the transcript of PDGFRA, and with normal PDGFRA nucleotide sequence comparing difference.

10. method as claimed in claim 9 is characterized in that, described difference is following single nucleotide polymorphism:

301 A → G;

Wherein, the nucleotide position numbering is based on SEQ ID NO:1.