CN111560428A

CN111560428A - Application of substance for detecting single nucleotide polymorphism of mitochondrial DNA rs3937033

Info

Publication number: CN111560428A
Application number: CN202010434691.1A
Authority: CN
Inventors: 王艳; 封志纯
Original assignee: 7th Medical Center of PLA General Hospital
Current assignee: 7th Medical Center of PLA General Hospital
Priority date: 2020-05-21
Filing date: 2020-05-21
Publication date: 2020-08-21
Anticipated expiration: 2040-05-21
Also published as: CN111560428B

Abstract

The invention discloses an application of a substance for detecting single nucleotide polymorphism of mitochondrial DNA rs 3937033. The invention provides a technical scheme for detecting the polymorphism or genotype of rs3937033 in a human genome and application of a substance for detecting the polymorphism or genotype of rs3937033 in preparation of products for detecting or assisting in detection of risk of high altitude pulmonary edema. rs3937033 can be used for screening individuals carrying human high altitude pulmonary edema susceptibility genes, predicting human susceptibility to high altitude pulmonary edema, screening individuals with high altitude pulmonary edema susceptibility and evaluating risk of high altitude pulmonary edema.

Description

Application of substance for detecting single nucleotide polymorphism of mitochondrial DNA rs3937033

Technical Field

The invention relates to application of a substance for detecting single nucleotide polymorphism, in particular to application of the substance for detecting single nucleotide polymorphism of mitochondrial DNARs3937033 in personalized health care detection service.

Background

Single Nucleotide Polymorphism (SNP) refers to a variation of a single nucleotide in a genome, which is the most minute variation unit and is a variation form formed by substitution, inversion, insertion or deletion of a single nucleotide pair. Single nucleotide polymorphisms are a high density of genetic markers on the genome, with more than 3000 tens of thousands of SNPs found in the human genome. As a third generation genetic marker, the SNP has a large number and dense distribution, and is easy to detect, so the SNP is an ideal genotyping target. SNP typing detection has great significance in the research of disease genome (such as disease susceptibility), drug genome (drug effect, drug metabolism difference and adverse reaction), population evolution and the like.

The organism mitochondria contain a plurality of genetic materials, namely mitochondrial DNA (mtDNA), and the mtDNA is an extracellular circular double-stranded DNA molecule and consists of an inner light chain (L) and an outer heavy chain (H). The total length of human mtDNA is 16,569bp, and the mtDNA consists of a coding region and a non-coding region, wherein the coding region has the functions of coding 13 polypeptides related to oxidative phosphorylation, 2 groups of rRNA and 22 tRNAs. Because mtDNA has no intron, and mtDNA is expressed as maternal inheritance, and no histone is combined to protect the coding process, the mtDNA is easy to mutate, and can influence the genetic information carried by some important segments in a genome, and finally, diseases are caused.

The mitochondrial DNA T16519C single nucleotide polymorphism (rs3937033) is a SNP site of a double-site polymorphism on human mitochondrial DNA, and the variation is a transition (T/C, A/G on its complementary strand). rs3937033 is located at MT: 16519(GRCh38), HGVS: NC-012920.1: m.16519T > C, located in MT-D-loop gene.

High Altitude Pulmonary Edema (HAPE) is the most common acute severe altitude disease in the plateau region, and if the treatment is not timely carried out, the life of the patient is endangered, and the problem is also a big problem which troubles the highland combat troops of various countries. In recent years, a series of studies have been made on the pathogenesis of HAPE at home and abroad, and unfortunately, the pathogenesis is still unclear, and the possible mechanisms which are currently accepted more generally are: on the one hand, the pulmonary vessels and alveolar respiratory epithelium of sensitive individuals have some important functional defects due to congenital genetic or acquired factors, and may cause excessive hypoxic pulmonary hypertension upon acute hypoxic exposure, which is the most critical link in HAPE development. On the other hand, impairment of the body's ability to clear the alveolar fluid due to genetic defects and impaired sodium transport from hypoxia to the alveolar epithelium may be a non-negligible important aspect of the promotion of HAPE.

Disclosure of Invention

The invention aims to solve the technical problem of how to screen individuals carrying non-susceptible genes of the high altitude pulmonary edema of a human or predict the susceptibility of the human to the high altitude pulmonary edema or screen susceptible individuals of the high altitude pulmonary edema or evaluate the risk of the high altitude pulmonary edema.

In order to solve the technical problem, the invention firstly provides any one of the following applications:

1. the application of the substance for detecting the polymorphism or genotype (namely allele) of rs3937033 in human genome in the preparation of products for detecting or assisting in detecting the risk of high altitude pulmonary edema.

2. Application of a substance for detecting the polymorphism or genotype (namely allele) of rs3937033 in a human genome in preparation of products for detecting or assisting in detection of the susceptibility of high altitude pulmonary edema.

3. Application of a substance for detecting rs3937033 polymorphism or genotype (namely allele) in human genome in preparation of products for screening or assisting in screening susceptible individuals or non-susceptible individuals of high altitude pulmonary edema.

4. The application of the substance for detecting the polymorphism or genotype (namely allele) of rs3937033 in the human genome in the preparation of products for evaluating or assisting in evaluating the risk of the high altitude pulmonary edema.

6. Application of a substance for detecting rs3937033 polymorphism or genotype (namely allele) in human genome in preparation of products for screening or assisting in screening of plateau pulmonary edema susceptible genotype or plateau pulmonary edema non-susceptible genotype.

In order to solve the technical problems, the invention also provides a product containing a substance for detecting the polymorphism or genotype (i.e. allele) of rs3937033 in the human genome.

The product containing the substance for detecting the polymorphism or genotype (namely allele) of rs3937033 in the human genome provided by the invention is any one of the products a) to d):

a) detecting a single nucleotide polymorphism or genotype product associated with high altitude pulmonary edema;

b) identifying or aiding in identifying a product of a single nucleotide polymorphism or genotype associated with high altitude pulmonary edema;

c) screening or assisting in screening products for patients with high altitude pulmonary edema;

d) detecting or assisting in detecting a product susceptible to high altitude pulmonary edema.

In the above, the detecting of the polymorphism or genotype of rs3937033 in human genomic mitochondrial DNA may specifically be detecting the nucleotide species of rs 3937033. The polymorphism or genotype of rs3937033 is T or C. The T is the genotype with the site of rs3937033 as the T, and the C is the genotype with the site of rs3937033 as the C. And the risk of developing high altitude pulmonary edema of the individual with the rs3937033 polymorphism or genotype C is lower than or is candidate for the individual with the rs3937033 polymorphism or genotype T.

Hereinbefore, the product may be suitable for Chinese, such as in plain regions. The Chinese people can be Han nationality people, and the gender can be male.

As used herein above, the substance for detecting the polymorphism or genotype (i.e., allele) of rs3937033 in human genomic mitochondrial DNA may be a reagent and/or an instrument required for determining the polymorphism or genotype of rs3937033 by at least one of the following methods: DNA sequencing, restriction enzyme fragment length polymorphism, single-strand conformation polymorphism, denaturing high performance liquid chromatography and SNP chip. The SNP chip comprises a chip based on nucleic acid hybridization reaction, a chip based on single base extension reaction, a chip based on allele-specific primer extension reaction, a chip based on one-step reaction, a chip based on primer connection reaction, a chip based on restriction enzyme reaction, a chip based on protein DNA binding reaction and a chip based on fluorescent molecule DNA binding reaction.

As described above, the substance for detecting the polymorphism or genotype of rs3937033 in human genomic mitochondrial DNA may contain a PCR primer and/or a single base extension primer for amplifying a human genomic DNA fragment including rs 3937033.

The sequence of the PCR primer has no special requirement, as long as the genomic DNA fragment including rs3937033 can be amplified, for example, the PCR primer can be a primer pair composed of single-stranded DNAs shown by SEQ ID No.1 and SEQ ID No.2 in the sequence table, or the PCR primer can also be a primer pair composed of single-stranded DNAs shown by sequence 4 and sequence 5 in the sequence table. The single base extension primer can be designed according to the upstream or downstream (excluding the SNP site) of rs3937033 in a human genome, the extended nucleotide of the single base extension primer corresponds to the nucleotide of the rs3937033 site of the human genome, namely, the 3' terminal nucleotide of the single base extension primer corresponds to the adjacent nucleotide of the rs3937033 of the human genome (namely, the former nucleotide of the rs3937033 or the latter nucleotide of the rs3937033), and for example, the single base extension primer can be specifically a single-stranded DNA shown as SEQ ID No.3 in a sequence table.

In the above, the product may be a reagent or a kit or a system, and the system may comprise a combination of a reagent or a kit and an apparatus, such as a product consisting of a PCR primer, a single base extension primer and a mass spectrometer, a combination of a PCR primer and a DNA sequencer, a combination of a PCR reagent and a DNA sequencing reagent and a DNA sequencer. The product can comprise the substance for detecting the polymorphism or the genotype of the rs3937033 in the mitochondrial DNA of the human genome.

In one embodiment of the invention, a genome DNA segment including mitochondrial DNA rs3937033 is amplified by using a PCR primer, a single base extension reaction is performed by using an obtained PCR amplification product as a template and using a single base extension primer, and the sequence of the obtained extension product is detected to determine the polymorphism and genotype of rs 3937033.

Experiments prove that in a case population consisting of 49 Chinese male Han nationality high pulmonary edema patients irrelevant on the blood margin in the plain region and a control population consisting of 58 Chinese male Han nationality normal people irrelevant on the blood margin in the plain region, the distribution frequency of the gene type of rs3937033 polymorphism sites of mitochondrial DNA in the case group is obviously lower than that in the control group (p-value is 0.001, and df is 1). Genotype C was found to reduce the probability of suffering from high altitude pulmonary edema, corrected by Logistic regression analysis (OR 0.259; 95% CI, 0.116-0.578; p-adjust ═ 0.002). The mitochondrial DNA rs3937033 is the single nucleotide polymorphism related to the high altitude pulmonary edema, the risk of HAPE occurrence of the population carrying the C allele at the site of T16519C (rs3937033) is obviously lower than that of the population carrying the T allele at the site of T16519C (rs3937033), and the rs3937033 can be used for screening individuals carrying the human high altitude pulmonary edema susceptibility genes, predicting the susceptibility of the human to the high altitude pulmonary edema, screening the individuals and evaluating the risk of the high altitude pulmonary edema. In practical application, in order to improve accuracy, the substance for detecting the rs3937033 polymorphism and genotype and other substances (such as substances for detecting other single nucleotide polymorphisms or genotypes related to the high altitude pulmonary edema) can be combined together to prepare a product for screening the individuals susceptible to the high altitude pulmonary edema.

Detailed Description

The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention. The examples provided below serve as a guide for further modifications by a person skilled in the art and do not constitute a limitation of the invention in any way.

The experimental procedures in the following examples, unless otherwise indicated, are conventional and are carried out according to the techniques or conditions described in the literature in the field or according to the instructions of the products.

Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

Example 1 analysis of polymorphic site rs3937033 of mitochondrial DNA and susceptibility to high altitude pulmonary edema

Statement of ethics

Each participant signed an informed consent form and the study was approved by the seventh medical center of the liberty military and the medical ethics committee of the general hospital of the tibetan liberty military.

Study object

The study objects are two populations obtained by the seventh medical center of the liberty military from 2018 to 2019 of the general hospital of the tibetan liberty military, wherein the population 1 consists of 49 patients with high altitude pulmonary edema (case group) and 58 normal persons (control group); population 2 (validation sample) peripheral blood samples from 33 patients with high altitude pulmonary edema (case group) and 19 normal persons (control group). In the two groups, 82 patients with high altitude pulmonary edema and 77 normal people are Han nations of Chinese male unrelated in blood margin in plain region. 82 patients with high altitude pulmonary edema aged 19-60 years, with the average (30.7 + -10.7) years old; 77 normal individuals were from a general physical examination population aged 19-55 years, with a mean (25.4 + -6.1) of years. All people had no history of smoking, drinking, and mitochondrial-related diseases such as cardiovascular disease, diabetes, etc., and had no history of previous altitude trips, and were all people who arrived at the same altitude tibetan pizza (3658 m).

In addition to meeting the above conditions, patients also need to meet the diagnosis standards of high altitude pulmonary edema, which are classified into on-site diagnosis standards and clinical diagnosis standards.

Criteria for on-site diagnosis

(1) The disease is developed. The recent arrival at the plateau (typically above 3000m altitude).

(2) Signs. Dyspnea at rest, chest tightness, oppression and feeling, cough, white or pink foamy sputum, weakness or reduced mobility.

(3) Signs. One or both lung fields show moist rales or wheezing, cyanosis medially, tachypnea, tachycardia.

At least two of the above symptoms and signs can be diagnosed.

Clinical diagnostic criteria

(1) The sputum reaches plateau (generally above an altitude of 3000 m) recently, and dyspnea, cough and white or pink foamy sputum appear at rest.

(2) Cyanosis medialis, wet rale of lung.

(3) The chest X-ray is the main basis of diagnosis, and the lung fields at one side or two sides of the lung with the lung portal as the center are in a spot-sheet shape or cloud-flocculent infiltration shadow which is often in diffuse and irregular distribution and can also be fused into a large-sheet shadow; the heart shadow is normal, but pulmonary hypertension and enlarged right heart can also be seen.

(4) Clinical examination and electrocardiogram examination can be performed to eliminate myocardial infarction, heart failure and other cardiopulmonary diseases, and pneumonia can be eliminated.

(5) After the treatment of bed rest, oxygen inhalation and the like or low-speed rotation, the symptoms are quickly improved, and the X-ray symptoms can disappear in a short time.

rs3937033 is a SNP site on human mitochondrial DNA, where the variation is a switch (T/C, A/G on its complementary strand). rs3937033 is located at MT: 16519(GRCh38), HGVS: NC-012920.1: m.16519T > C, located in MT-D-loop gene (NCBI Reference Sequence: NC-012920.1, 31-OCT-2014). The rs3937033 genotype is T or C. Wherein T is the genotype with the site of rs3937033 as T, and C is the genotype with the site of rs3937033 as C.

1. Extraction of genomic DNA

Genomic DNA of each individual was extracted and used as a template for genotype detection.

2. Design and Synthesis of primers

The PCR amplification primer of rs3937033 and the single-base extension primer were designed using the Genotyping Tools of Sequenom and MassARRAY Assay Design software, and were synthesized by Biotech. The sequence of the PCR amplification specific primer pair for detecting the rs3937033 locus genotype of the mitochondrial DNA gene is as follows: a forward primer: 5'-ACGTTGGATGCCATAACACTTGGGGGTAG-3' (SEQ ID No.1 of the sequence Listing); reverse primer: 5'-ACGTTGGATGCGTGATGTCTTATTTAAGGGG-3' (SEQ ID No.2 of the sequence Listing). The single base extension primer sequence for the single base amplification reaction is: 5'-CTGGTTCCTACTTCAGGG-3' (SEQ ID No.3 of the sequence Listing)

3. Establishment of method for detecting genotype of mitochondrial DNA rs39370334 site

3.1PCR amplification

PCR amplification was performed in 384-well plates, and the total volume of each reaction system, excluding the template, was 4. mu.L, and PCR reaction systems were prepared as shown in Table 1.

TABLE 1 Components of each PCR reaction System

Reagent	Volume (μ L)
		10 × PCR buffer	0.5
MgCl₂(25mM)	0.4
		dNTP mix(25mM)	0.1
HotStar Taq enzyme (5U/. mu.L)	0.1
		Ultrapure water	1.9
DNA shown in sequence 1	0.5
		DNA shown in sequence 2	0.5
Total volume	4

The genomic DNA sample prepared in 1 above was removed, and the sample addition volume was adjusted to 1. mu.L, and each 5. mu.L PCR reaction system contained 20-50ng of template DNA, 0.5U of Hotstar Tag, 0.5pmol of each amplification primer, and 0.1. mu.L of 25mM dNTPs.

The PCR reaction program is: 4 minutes at 94 ℃; 94 ℃ for 20 seconds, 56 ℃ for 30 seconds, 72 ℃ for 1 minute, 45 cycles; 3 minutes at 72 ℃; keeping at 4 ℃.

Obtaining PCR products.

3.2 alkaline phosphatase treatment of PCR products

After completion of the PCR reaction, the PCR product obtained in 3.1 above was treated with SAP (shrimp alkaline phosphatase) to remove free dNTPs from the system, and an SAP reaction system was prepared as shown in Table 2.

TABLE 2 SAP reaction System

Reagent	Volume (μ L)
		Ultrapure water	1.53
10 × SAP buffer	0.17
		SAP enzyme (1.7U/ul)	0.3
PCR product	5
		Total volume	7

The reaction conditions are as follows: 40 minutes at 37 ℃; 5 minutes at 85 ℃; maintaining the temperature at 4 ℃.

The alkaline phosphatase-treated product was obtained.

3.3 Single base extension

After the alkaline phosphatase treatment, the single-base extension reaction was carried out, and a single-base extension reaction system was prepared as shown in Table 3.

TABLE 3 Single-base extension reaction System

Reagent	For each reaction, μ L
		Water (W)	1.53
10 × buffer for single-base extension reaction	0.17
		Single base extension reaction enzyme (1.7U/ul)	0.3
Alkaline phosphatase treated product	7
		DNA molecule shown in sequence 3	1
Total volume	10

The reaction conditions are as follows:

i.94 ℃ for 30 seconds

II.94 ℃ for 5 seconds

III.52 ℃ for 5 seconds

IV.80 ℃ for 5 seconds

V. Return to III, 4 cycles

VI, Return to II, 39 cycles

VII.72 ℃ for 3 minutes

VIII.4℃

Obtaining the single base extension product.

3.4 resin purification

Clean Resin (Sequenom, USA) was spread into a 6mg Resin plate; adding 16 μ l of water to the corresponding well of the single base extension product obtained in 3.3 above; pouring the dried resin into an extension product plate, sealing the film, and vertically rotating at a low speed for 30 minutes to ensure that the resin is fully contacted with reactants; the resin was allowed to settle to the bottom of the well by centrifugation to give a resin purified extension product.

3.5 chip spotting

The MassARRAY Nanodispenser RS1000 Spotter (SEQUENOM) was started and the resin-purified extension product was transferred to a 384-well SpectroCHIP (Sequenom) chip (SEQUENOM).

3.6 Mass spectrometric detection

The spotted SpectroCHIP chip is analyzed by MALDI-TOF, and the detection result is typed by TYPER 4.0 software (sequenom) and output.

4. The determination of the mitochondrial DNA polymorphic site rs3937033 genotype and the analysis of the susceptibility of the mitochondrial DNA polymorphic site and the high altitude pulmonary edema respectively extract the genomic DNA of the peripheral blood samples of 49 high altitude pulmonary edema patients and a first group of 58 normal persons in the population 1, and carry out PCR amplification, single base extension reaction and genotyping. The distribution of genotype frequencies of the rs3937033 polymorphic sites of the two sets of mitochondrial DNA is shown in table 4. 17 individuals with the rs3937033 genotype C in 49 patients with high altitude pulmonary edema have the genotype frequency of 34.7 percent; 32 individuals with the rs3937033 genotype T have the genotype frequency of 65.3%. 39 individuals with the rs3937033 genotype C in 58 normal people have the genotype frequency of 67.2 percent; 19 individuals with the rs3937033 genotype T have the genotype frequency of 32.8 percent. All data were processed using an independent sample Person test with SPSS17.0(SPSS inc., USA) statistical software.

In all samples, the distribution frequency of C genotype in case group (49 patients with high pulmonary edema) was significantly lower than that in control group (p-value 0.001, df 1) compared with that in control group (58 normal persons), indicating that there was a significant difference in distribution frequency of C, T genotype between case group and control group (p < 0.05). An increase in the C genotype was found to decrease the probability of HAPE after correction by Logistic regression analysis (OR 0.259; 95% CI, 0.116-0.578; p-adjust ═ 0.002). The results show that the mitochondrial DNA polymorphic site rs3937033 is significantly related to the high altitude pulmonary edema. The risk of HAPE in the population carrying the T16519C (rs3937033) locus C allele was significantly lower than in the population carrying the T16519C (rs3937033) locus T allele.

TABLE 4 genotype frequencies of the rs3937033 polymorphic sites of mitochondrial DNA of case and control groups

Note: p-value: person' s²test, p-adjust: adjust p-value, OR: ratio of ratios, CI: a confidence interval.

5. Verification of mitochondrial DNA polymorphic site rs3937033 genotype

The genomic DNA from the peripheral blood samples of 33 patients with high altitude pulmonary edema and 19 normal persons from population 2 were again selected according to the same principle and method, and the DNA was amplified using conventional PCR, forward primer: 5'-CAGCCACCATGAATATTGTACG-3' (SEQ ID No.4 of the sequence Listing); reverse primer: 5'-GTTAGGCTGGTGTTAGGGTTC-3' (SEQ ID No.5 in the sequence table), amplifying a genome DNA fragment including the rs3937033 site of the mitochondrial DNA gene by PCR and sequencing, wherein the result shows that two PCR products are obtained together, the sequences of the two PCR products are the sequence 6 in the sequence table, and in the sequence 6, y is c or t. The rs3937033 site is positioned at the 412 th site of the sequence 6, the nucleotide is C or T, when the nucleotide is T, the rs3937033 genotype is T, and when the nucleotide is C, the rs3937033 genotype is C. The distribution of the genotype frequencies of the rs3937033 polymorphic sites of the two sets of mitochondrial DNAs is shown in Table 5. The result shows that 7 individuals with the rs3937033 genotype as C in 33 patients with high altitude pulmonary edema have the genotype frequency of 21.2 percent; 26 individuals with the rs3937033 genotype T have the genotype frequency of 78.8%. 10 individuals with the rs3937033 genotype as C in 19 normal people have the genotype frequency of 52.6 percent; 9 individuals with the rs3937033 genotype T have the genotype frequency of 47.4%. Data were processed for statistics using an independent sample Person test of SPSS17.0(SPSS inc., USA) statistical software.

TABLE 5 genotype frequencies of the rs3937033 polymorphic sites of mitochondrial DNA of case and control groups

In all the samples tested, the distribution frequency of C genotype in case group (33 patients with high pulmonary edema) was significantly lower than that in control group (p-value of 0.020 and df of 1) compared with that in control group (19 normal persons), indicating that there was a significant difference in the distribution frequency of C, T genotype between case group and control group (p < 0.05). An increase in the C genotype was found to decrease the probability of HAPE after correction by Logistic regression analysis (OR 0.242; 95% CI, 0.071-0.827; p-adjust ═ 0.044). The results show that the mitochondrial DNA polymorphic site rs3937033 is significantly related to the high altitude pulmonary edema. Consistent with the results in table 4.

All the results prove that the risk of HAPE of the population carrying the C allele at the T16519C (rs3937033) locus is obviously lower than that of the population carrying the T allele at the T16519C (rs3937033), and the rs3937033 can be used for screening individuals susceptible to high altitude pulmonary edema, predicting the susceptibility of people to high altitude pulmonary edema, screening individuals carrying the genes susceptible to high altitude pulmonary edema and evaluating the risk of high altitude pulmonary edema.

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Claims

1. The application of the substance for detecting the polymorphism or genotype of rs3937033 in the human genome in the preparation of products for detecting or assisting in detecting the risk of the high altitude pulmonary edema.

2. The application of the substance for detecting the polymorphism or genotype of rs3937033 in human genome in preparation of products for detecting or assisting in detection of susceptibility to high altitude pulmonary edema.

3. The application of the substance for detecting the polymorphism or genotype of rs3937033 in the human genome in preparing products for screening or assisting in screening susceptible individuals or non-susceptible individuals of high altitude pulmonary edema.

4. The application of the substance for detecting the polymorphism or genotype of rs3937033 in the human genome in the preparation of products for evaluating or assisting in evaluating the risk of the disease of the high altitude pulmonary edema.

5. Use according to any one of claims 1 to 4, characterized in that: the polymorphism or genotype of the rs3937033 is T or C, the T is the polymorphism or genotype of the site rs3937033 as T, and the C is the polymorphism or genotype of the site rs3937033 as C; and the risk of developing high altitude pulmonary edema of the individual with the rs3937033 polymorphism or genotype C is lower than or is candidate for the individual with the rs3937033 polymorphism or genotype T.

6. The application of the substance for detecting the polymorphism or genotype of rs3937033 in a human genome in preparing products for screening or assisting in screening the susceptible genotype or the non-susceptible genotype of plateau pulmonary edema.

7. Use according to claim 6, characterized in that: the susceptible genotype of the plateau pulmonary edema is T, and the non-susceptible genotype of the plateau pulmonary edema is C; the T is the genotype with the site of rs3937033 as the T, and the C is the genotype with the site of rs3937033 as the C.

8. Use according to any one of claims 1 to 7, characterized in that: the substance for detecting the polymorphism or genotype of the rs3937033 in the human genome contains a PCR primer and/or a single-base extension primer for amplifying a human genome DNA fragment including the rs 3937033.

9. A product containing a substance for detecting the polymorphism or genotype of rs3937033 in human genome, which is any one of a) to d):

10. The product of claim 9, wherein: the substance for detecting the polymorphism or genotype of the rs3937033 in the human genome contains a PCR primer and/or a single-base extension primer for amplifying a human genome DNA fragment including the rs 3937033.