CN101294199A - Coronary heart disease detection method and kit - Google Patents

Abstract

The invention discloses a method for detecting the susceptibility of atherosclerosis and/or coronary heart disease, which comprises detecting whether there is variation in individual vascular cell adhesion molecule 1 (VCAM1), transcript and/or protein compared with normal, The presence of a variant 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 concerning the single nucleotide polymorphism (SNP) of vascular cell adhesion molecule type-1 (VCAM1) and its association with atherosclerosis and/or 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.

For a long time, research at home and abroad has 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 VCAM1 gene and atherosclerosis and/or coronary heart disease, let alone the confirmation of VCAM1 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 VCAM1 gene, transcript and/or protein, and comparing it with normal VCAM1 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 VCAM1, and compares the difference with the normal VCAM1 nucleotide sequence.

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

256-bit C→T;

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 vascular cell adhesion molecule 1 (VCAM1) exists in a sample, comprising the steps of:

(a) amplifying the VCAM1 gene of the sample with VCAM1 gene-specific primers to obtain an amplification product; and

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

256-bit C→T;

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 256th position in SEQ ID NO:1.

In a third aspect of the present invention, a test kit for detecting atherosclerosis and/or coronary heart disease is provided, which includes primers for specifically amplifying the VCAM1 gene or transcripts, more preferably, the primers amplify The length is 100-2000bp, and contains the amplification product at position 256 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 256 in SEQ ID NO: 1;

(b) Recognizing the mutant restriction enzyme at position 256 in SEQ ID NO:1.

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

256-bit C→T;

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 VCAM1 is closely related to atherosclerosis and/or coronary heart disease. The results of the association study showed that the SNP at position 256 in SEQ ID NO: 1 of VCAM1 (position 256 C→T)( Denoted as RS3176867) there is a significant difference in distribution between cases and controls (P<0.05), 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.

VCAM1 gene

The sequence of vascular cell adhesion molecule type-1 (VCAM1) is known, and its detailed sequence and some related information can be found at http://www.ncbi.nlm.nih.gov/Genebank/; or http://www.ncbi.nlm.nih.gov/SNP. For convenience, the nucleotide sequence relevant to the SNP of the present invention in VCAM1 is given in SEQ ID NO: 1.

The VCAM1 gene is a support protein of vascular epithelial cells and plays a role in signal transduction. This gene is considered to play an important role in the early stage of atherosclerosis. The loss of LOH of VCAM1 gene is one of the reasons for inducing vascular sclerosis (Arvanitis DA et al. J Cell Mol Med. 2005; 9(1): 153-9).

The inventors sequenced almost the entire region of the VCAM1 gene and found many SNPs, most of which were not associated with atherosclerosis and/or coronary heart disease susceptibility, but association studies showed that 256 in SEQID NO:1 However, the position C→T is a SNP highly correlated with susceptibility to atherosclerosis and/or coronary heart disease.

Specifically, the present invention discloses the single nucleotide polymorphism (SNP) of No. 4 intron of VCAM1 gene and the correlation between the polymorphism and coronary atherosclerosis. The SNP of the present invention is the C/T polymorphism of the 256th position of the sequence shown in SEQ ID NO: 1, which is located in the No. 4 intron of the human VCAM1 gene, and the homozygous TT in the coronary atherosclerosis patient population The frequency was significantly higher than that in the normal control population.

Based on the new discovery of the present invention, the VCAM1 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 VCAM1 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 VCAM1 DNA or polypeptides encoded by its fragments, especially monoclonal antibodies. Here, "specificity" means that the antibody can bind to human VCAM1 gene product or fragment. Preferably, it refers to those antibodies that can bind to human VCAM1 gene products or fragments but do not recognize and bind to other irrelevant antigen molecules. Antibodies in the present invention include those molecules capable of binding and inhibiting human VCAM1 protein, as well as those antibodies that do not affect the function of human VCAM1 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, purified human VCAM1 gene product, or antigenic fragments thereof, can be administered to animals to induce polyclonal antibody production. Similarly, cells expressing human VCAM1 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 antibody of the present invention includes the antibody capable of blocking the function of human VCAM1 protein and the antibody not affecting the function of human VCAM1 protein. All kinds of antibodies of the present invention can be obtained by conventional immunization techniques by utilizing fragments or functional regions of human VCAM1 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 VCAM1 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 VCAM1 protein can be used in immunohistochemical techniques to detect the amount and/or mutation of human VCAM1 protein in biopsy specimens. A preferred anti-VCAM1 antibody is an antibody that does not recognize normal VCAM1 but recognizes mutant VCAM1, or an antibody that recognizes normal VCAM1 but not mutant VCAM1. Using these antibodies, the detection of atherosclerosis and/or coronary heart disease susceptibility at the protein level can be conveniently performed.

Using the VCAM1 protein of the present invention, substances that interact with the VCAM1 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 VCAM1 protein level. These assays are well known in the art and include ELISA and the like.

A method for detecting whether there is VCAM1 protein in a sample is to use a specific antibody for VCAM1 protein to detect, which includes: contacting the sample with a specific antibody for VCAM1 protein; observing whether an antibody complex is formed, which means that the antibody complex is formed VCAM1 protein is present in the sample.

The polynucleotide of VCAM1 protein can be used for auxiliary diagnosis of VCAM1 protein-related diseases. In terms of diagnosis, the polynucleotide of VCAM1 protein can be used to detect the expression of VCAM1 protein or the abnormal expression of VCAM1 protein in a disease state. For example, VCAM1 DNA sequence can be used for hybridization of biopsy specimens to determine the abnormal expression of VCAM1 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 VCAM1 protein-specific primers can also detect the transcripts of VCAM1 protein.

Detection can be against cDNA or genomic DNA. The forms of VCAM1 protein mutations include point mutations, translocations, deletions, recombinations, and any other abnormalities compared with the normal wild-type VCAM1 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 VCAM1 gene of the sample with VCAM1 gene-specific primers to obtain the amplified product; then detect whether the following single nucleotide polymorphism exists in the amplified product: 256 C → T, wherein the nucleotide position numbering is based on SEQ ID NO:1.

It should be understood that after the present invention reveals for the first time the correlation between the SNP of the VCAM1 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 of the SNP. The product, and then determine whether there is 256 C→T 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 256th 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.

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 187 patients and 187 normal controls 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 VCAM1 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 gggcattata tctaattaga agaca 2 antisense primer aaattgaata gataaaaatt tgtca 3

1.2.3 PCR amplification of VCAM1 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 VCAM1 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: C→T at position 256 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 187 patients and 187 normal controls, the 256 SNPs of VCAM1 were significantly different between the patient group and the control group: the frequency of homozygous TT was 4.3% in the patient population and 4.3% in the control group. The average value was 1.1%, and there was a significant difference between the two in the 90% confidence interval (p=0.070).

The above results confirm that the SNP at position 256 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 C→T at position 256 in SEQ ID NO: 1 is closely related to atherosclerosis and/or coronary heart disease. Therefore, based on this mutation, VCAM1 gene-specific primers can be designed to amplify and detect with the patient's DNA as a template.

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

102 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.

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 the 256-position C→T can also be detected.

Test results:

For subjects with 256 positions C→T in VCAM1, it is further detected by conventional methods to confirm whether they suffer from atherosclerosis. The test results show that the susceptibility ratio of atherosclerosis and/or coronary heart disease of the subjects containing 256 C→T (especially the TT purifier) is significantly higher than that of the normal population with genotype CC or CT (at least 3 times).

Therefore, this indicates that auxiliary detection and/or early diagnosis of atherosclerosis can be performed by detecting 256 C→T of VCAM1.

Example 3

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

The test in Example 2 was repeated, with the difference that 90 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.

As a result, 6 subjects were detected to be TT homozygotes at the SNP site of the present invention, and further confirmed by conventional arteriography, among which 4 subjects had atherosclerosis.

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.

sequence listing

<110>Shanghai Human Genome Research Center

<120> Coronary heart disease detection methods and kits

<130>072052

<160>3

<170>PatentIn version 3.1

<210>1

<211>511

<212>DNA

<213> Homo sapiens

<400>1

gggcattata tctaattaga agacattttt tgtagctttt tccagtacct cttttagacc 60

attttatcac catatgagga tgacttgatt tagttgttgt agcaactgaa aagctgggat 120

cttgaatcag accccacctgg catcaagtcc taattttgct acttgcaaac tgggcaaagt 180

tatttagcat ttctgagcct aaatttcctc aagaagaaaa aggaggggga aaaggaggaa 240

gagaaagagt agaagcagaa agagaaggca aagaagaaag aatagtacta gtgtacaaaa 300

ttgttgtgag gattaaatga gaatatatat atatatatat atatatatat atatagtaat 360

ttagatagtg cttgacacac aagaaccact taaattattt gagtgattgt ccactcttac 420

agtgatttga taaactacat ttgagcagat actctattat tgctgtgaag agatagcaac 480

ctctcttgac aaatttttat ctattcaatt t 511

<210>2

<211>25

<212>DNA

<213> Artificial sequence

<220>

<221>misc_feature

<223> Primer

<400>2

gggcattata tctaattaga agaca 25

<210>3

<211>25

<212>DNA

<213> Artificial sequence

<220>

<221>misc_feature

<223> Primer

<400>3

aaattgaata gataaaaatt tgtca 25

Claims (10)

1. a method for in vitro detection of whether there is a single nucleotide polymorphism of vascular cell adhesion molecule 1 (VCAM1), characterized in that, comprising steps: (a) amplifying the VCAM1 gene of the sample with VCAM1 gene-specific primers to obtain an amplification product; and (b) Detect whether the following single nucleotide polymorphisms exist in the amplification product: 256-bit C→T; Wherein, the nucleotide position numbers are based on SEQ ID NO:1. 2. The method of claim 1, wherein the gene-specific primer has the sequences of SEQ ID NO: 2 and 3. 3. The method according to claim 1, wherein the length of the amplified product is 100-2000bp and contains the 256th position in SEQ ID NO:1. 4. A test kit for detecting atherosclerosis and/or coronary heart disease, characterized in that it includes primers for specific amplification of the VCAM1 gene or transcripts, and the amplified primers are 100-2000bp in length and contain The amplified product at position 256 in SEQ ID NO:1. 5. The kit of claim 4, further comprising reagents selected from the group consisting of: (a) a probe that binds to the mutation at position 256 in SEQ ID NO: 1; (b) Recognizing the mutant restriction enzyme at position 256 in SEQ ID NO:1. 6. test kit as claimed in claim 5, is characterized in that, described mutation is following single nucleotide polymorphism: 256-bit C→T; Wherein, the nucleotide position numbers are 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. A method for diagnosing individual susceptibility to atherosclerosis and/or coronary heart disease, characterized in that it comprises the steps of: Detecting the individual's VCAM1 gene, transcript and/or protein, and comparing it with normal VCAM1 gene, transcript and/or protein, Wherein, the existence of a difference indicates that the possibility of the individual suffering from atherosclerosis and/or coronary heart disease is higher than that of the normal population. 9. The method according to claim 8, characterized in that the gene or transcript of VCAM1 is detected, and the difference is compared with the normal VCAM1 nucleotide sequence. 10. The method of claim 9, wherein said difference is the following single nucleotide polymorphism: 256-bit C→T; Wherein, the nucleotide position numbers are based on SEQ ID NO:1.