A breast cancer detection kit

Abstract

The present invention discloses a breast cancer detection kit, which includes a reagent for detecting the genotype of a TCHH gene susceptibility SNP site (1) and a susceptibility SNP site (2) in a sample. The present invention studies the application prospects of SNPs in breast cancer detection, describes the influence of SNPs on breast cancer progression, and reveals their diagnostic value. Therefore, the present invention, through the development and application of a SNP genotype detection kit, can make the diagnosis of breast cancer more convenient and easy, help clinicians quickly and accurately understand the patient's condition, lay the foundation for clinical treatment effect evaluation, and provide assistance for discovering new small molecule drug targets with potential therapeutic value.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to a breast cancer detection kit. Background Art

[0002] Breast cancer is a systemic disease whose occurrence and progression is a complex process involving multiple factors and multiple links, including the activation of oncogenes and the inactivation of tumor suppressor genes. Therefore, gene mutations play a very important role in the occurrence and development of breast cancer.

[0003] Breast cancer is a multifactorial disease with genetic variability, with less than 10% of cases caused by single gene defects. With the development of high-throughput genetic technologies, more and more breast cancer-related genes have been discovered. Potential genetic variations (single nucleotide polymorphisms and copy number variations) in these genes may contribute to differences in the efficacy of breast cancer drug treatments. These genetic variations may affect the metabolic pathways of anti-cancer drugs and the target genes of their action, thereby affecting efficacy and prognosis.

[0004] SNPs (single nucleotide polymorphisms) are a type of molecular genetic marker proposed in 1996 by Lander, a scholar at the Human Genome Research Center at the Massachusetts Institute of Technology. They primarily refer to DNA sequence polymorphisms caused by variations in a single nucleotide at the genomic level. SNPs represent polymorphisms involving variations in only a single base, manifesting as transitions, transversions, insertions, and deletions. SNPs are third-generation genetic markers, and many phenotypic differences in the human body, including susceptibility to drugs or diseases, may be associated with SNPs. Currently, predictive research on the prognosis and efficacy of different breast cancer subtypes focuses primarily on the SNP level.

[0005] SNPs confer distinct responses to environmental exposures, drug treatments, and other factors, resulting in distinct phenotypes. Therefore, SNPs may be an important genetic basis for individual differences in disease development and progression. Using SNP profiles to diagnose disease susceptibility is rapid, sensitive, and accurate, and thus holds great promise. In recent years, the use of SNPs to diagnose disease development and progression has become a hot topic among both clinical and scientific researchers.

[0006] However, there are currently no reports on the application of SNPs in breast cancer diagnosis. If SNPs that are susceptible to breast cancer can be screened out as biomarkers and corresponding diagnostic kits can be developed, it will surely greatly promote the current status of early diagnosis of breast cancer in my country and open up new avenues for its drug screening, efficacy evaluation and targeted treatment. Summary of the Invention

[0007] The purpose of the present invention is to solve the above technical problems and provide a breast cancer detection kit.

[0008] The inventors isolated and studied single nucleotide polymorphisms in the peripheral blood DNA of breast cancer patients and age-matched healthy female controls, searching for a group of highly specific and sensitive SNPs that are highly correlated with breast cancer. They also developed a breast cancer prognosis detection kit that can be easily used in clinical practice, providing data support for breast cancer detection.

[0009] The purpose of the present invention is achieved through the following technical solutions:

[0010] A breast cancer detection kit comprises a reagent for detecting the genotype of a TCHH gene susceptible SNP site (1) and a susceptible SNP site (2) in a sample.

[0011] The specific information of the TCHH gene susceptible SNP site (1) is rs181128140, NM_007113: exon3: c.G4646C: p.R1549P; the specific information of the TCHH gene susceptible SNP site (2) is NM_007113: exon3: c.G4678A: p.E1560K. The two susceptible SNP sites provided by the present invention are both located on the NM_007113 transcript of human chromosome 1, wherein the susceptible SNP site (1) is a missense mutation of the SNP site with a base change from G to C at position 152081047, and the susceptible SNP site (2) is a missense mutation of the SNP site with a base change from G to A at position 152081015.

[0012] Among them, TCHH (trichohyalin), according to information from the NCBI RefSeq database, encodes a protein that forms a cross-linked complex. This cross-linked complex, along with keratin intermediate filaments, provides mechanical support to the inner root sheath of the hair follicle. This encoded protein also plays an important role in maintaining the structural integrity of the filiform papilla of the tongue. Defects in this gene are the cause of unfusing hair syndrome. (Source, February 2017)

[0013] In addition, gene ontology analysis provided by UniProt-GOA showed that this gene is involved in many biological processes such as keratinization and has calcium ion binding function.

[0014] Furthermore, the present invention also provides a kit comprising two susceptible SNP sites respectively.

[0015] Preferably, the kit can be a reagent for detecting SNPs using any technique known in the art, as long as it can detect whether there is an allele mutation at the susceptible SNP site (1) and / or the susceptible SNP site (2) in a sample. It includes but is not limited to the embodiments listed below.

[0016] In a first embodiment, the kit includes reagents for detecting the presence of the C allele at the susceptible SNP site (1) and / or the A allele at the susceptible SNP site (2) in a sample using a sequencing method. Sequencing methods are well known in the art, and the required primers and other reagents can be selected by a person of ordinary skill in the art as needed (see the instructions for use of sequencers from companies such as ABI and Beckman), and will not be described in detail here. Using the kit, the sequence of the susceptible SNP site (1) and / or the susceptible SNP site (2) in the sample can be directly measured by sequencing, thereby determining whether the sample carries a variation in the corresponding site allele, and further determining the susceptibility to obesity.

[0017] In a second embodiment, the kit includes reagents for detecting the genotype of the susceptible SNP site (1) and / or the susceptible SNP site (2) in a sample using a Taqman probe SNP detection method. The Taqman probe used is a probe designed for the susceptible SNP site (1) and / or the susceptible SNP site (2). The probe can be provided by a reagent company or designed by the user using software such as Beacon Designer 7.5 from PREMIER Biosoft.

[0018] In a third embodiment, the kit is a kit for detecting the genotype of susceptible SNP sites (1) and / or susceptible SNP sites (2) in a sample using PCR-single-strand conformation polymorphism. The kit includes primers for amplifying susceptible SNP sites (1) and / or susceptible SNP sites (2), PCR reagents, control samples and reagents required for electrophoresis to detect conformation. The electrophoresis is preferably non-denaturing polyacrylamide gel electrophoresis. The control samples include negative or positive pure and control samples, and may also include or not include heterozygote corresponding heterozygote control samples. Preferably, the above three types of control samples are included at the same time. The amplified product of the sample to be tested and the amplified product of the control sample are electrophoresed simultaneously, and the comparison of the electrophoresis results can obtain the detection result of whether the sample to be tested carries the corresponding allele variation.

[0019] The kits of the first, second and third embodiments described above include primers designed for amplifying susceptible SNP sites (1) and / or susceptible SNP sites (2), and the primers have the following characteristics: 1) the length of the amplified product is between 100-350 bp; 2) the primer length is between 15 and 30 bases; 3) the G+C content is between 40% and 60%; 4) the bases are randomly distributed; 5) the primer itself cannot have four consecutive bases of complementarity; and 6) there cannot be four consecutive bases of complementarity between primers.

[0020] The primers can be designed using software (e.g., using Primer5, Oligo6, etc.). For example, preferably, the present invention uses a primer pair as shown in SEQ ID NO: 1-2 to simultaneously detect the genotypes of two susceptible loci. In addition, the present invention provides the use of the primer pair in preparing a breast cancer detection kit. If the presence of the susceptible SNP site (1) carrying the C allele and the susceptible SNP site (2) carrying the A allele is detected in the sample, it indicates that the individual providing the sample has a poor prognosis for breast cancer. The prepared kit can be at least one of the above kits, or can be other kits that use other techniques known in the art to identify the presence of the SNP.

[0021] Beneficial effects of the present invention:

[0022] This study investigates the potential application of SNPs in breast cancer detection, explains their impact on breast cancer progression, and reveals their diagnostic value. Therefore, through the development and application of a SNP genotyping kit, this study can make breast cancer diagnosis more convenient and accessible, enabling clinicians to quickly and accurately assess a patient's condition, laying the foundation for evaluating clinical treatment outcomes, and aiding the discovery of new small molecule drug targets with potential therapeutic value. DETAILED DESCRIPTION

[0023] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.

[0024] The technical solution of the present invention specifically includes: collecting blood samples that meet the standards and systematically collecting complete demographic and clinical data; genotype detection: selecting breast cancer cases and healthy female controls aged matched with the breast cancer cases, and using exome sequencing to identify SNPs associated with breast cancer; further using genotyping to detect the positively associated SNPs screened out to verify their repeatability in clinical diagnosis; and developing a breast cancer auxiliary diagnosis kit: developing a SNP auxiliary diagnosis kit based on SNPs with significantly different genotype distribution frequencies between breast cancer cases and healthy female controls.

[0025] The numerical values in the data analysis are expressed as follows:

[0026] 1. ljb23_sift: SIFT score (version 2.3), indicating the effect of the variant on the protein sequence. It contains three values: the initial SIFT score, the converted value (1-SIFT), and T or D. When the variant affects multiple protein sequences simultaneously, a SIFT score is assigned to each protein sequence, and the minimum value is taken. The smaller the SIFT score, the more "detrimental" it is, indicating that the SNP is likely to cause changes in protein structure or function; D: Deleterious (sift <= 0.05); T: Tolerated (sift > 0.05);

[0027] 2. ljb23_pp2hvar: PolyPhen2 is used to predict the effect of this variant on the protein sequence based on the HumanVar database, which is used for single-gene genetic diseases. This column contains two values: the first is the PolyPhen2 score, where a larger value indicates a greater likelihood of "damage" (D), indicating that the SNP may cause changes in protein structure or function; the second is D, P, or B (D: Probably damaging (>= 0.909), P: Possibly damaging (0.447 <= pp2_hvar <= 0.909); B: Benign (pp2_hvar <= 0.446)).

[0028] 3. ljb23_pp2hdiv: Uses PolyPhen2 to predict the effect of this variant on protein sequence based on the HumanDiv database, which is used for complex diseases. This column contains two values: the first is the PolyPhen 2 score, where a larger value indicates a greater likelihood of "damage" (D), P, or B (D: Probably damaging (>= 0.957), P: Possibly damaging (0.453 <= pp2_hdiv <= 0.956); B: Benign (pp2_hdiv <= 0.452)).

[0029] 4. ljb23_mt:tionTaster score (version 2.3), which indicates the effect of the variant on the protein sequence. It contains three values: the initial Mutation Taster score, the converted value, and A, D, N, or P. The larger the second value, the more "harmful", indicating that the SNP is more likely to cause changes in protein structure or function, including "A" ("disease causing automatic"), "D" ("disease causing"), "N" ("polymorphism"), and "P" ("polymorphism automatic").

[0030] Specifically, the experimental methods of the study mainly include the following parts:

[0031] 1. Selection of research samples

[0032] (1) 25 breast cancer cases diagnosed by pathology and 10 healthy women matched with the breast cancer cases as controls, 3 of the breast cancer cases had a family history of cancer;

[0033] (2) had not received radiotherapy or chemotherapy and had no history of tumor before blood collection;

[0034] (3) Healthy female controls matched with the cases in age

[0035] 2. Extract genomic DNA from peripheral blood using the phenol-chloroform method according to conventional procedures. Typically, 20-50 ng / μL DNA is obtained, with a purity (UV 2600D: 2800D) of 1.6-2.0.

[0036] 3. Whole-exome chip detection

[0037] (1) Obtain whole genome DNA samples from the subjects;

[0038] (2) Scanning on a whole exon chip (Beijing Novogene Technology Co., Ltd., the same below);

[0039] (3) Detect and compare the differences of each genotype in breast cancer cases and healthy female controls.

[0040] 4. Single SNP Genotyping

[0041] (1) Obtain DNA samples from subjects;

[0042] (2) Design specific amplification primers for a single SNP;

[0043] (3) Perform PCR reaction and recover the product for sequencing;

[0044] (4) Compare the distribution differences of different genotypes in breast cancer cases and healthy female controls.

[0045] 5. Preparation of diagnostic kit

[0046] Exome-wide microarray scanning and single SNP detection identify SNPs with significant differences in genotype distribution frequency between breast cancer cases and healthy female controls, which serve as diagnostic indicators for breast cancer. The diagnostic kit for SNPs identified as associated with breast cancer prognosis includes reagents for detecting the genotypes of two susceptible SNPs located in the TCHH gene. The kit may also include specific amplification primers for these SNPs, as well as Taq enzyme, dNTPs, and other reagents.

[0047] 6. Clinical Application Examples

[0048] The breast cancer detection kit developed by the inventors was used to test breast cancer patients and compared with actual clinical testing to confirm its effectiveness. Specifically, the kit includes specific amplification primers and other detection reagents for detecting the aforementioned SNPs in cDNA from blood samples of subjects. This allows clinicians to quickly and accurately assess a patient's disease status and severity, enabling them to implement more personalized prevention and treatment plans in a timely manner.

[0049] Example 1: Sample collection and sample data organization

[0050] The inventors collected a large number of blood samples from newly diagnosed breast cancer patients at Shenzhen Second People's Hospital between January 2010 and December 2015. After sorting the sample data, the inventors selected 25 samples that met the following criteria. They also selected 10 healthy women aged 25-55 as controls for whole-exome microarray testing. The sample selection criteria were as follows:

[0051] 1. Cases of breast cancer diagnosed by pathology, including 3 patients with a family history of cancer, labeled X1, X2, and X3;

[0052] 2. No radiotherapy or chemotherapy before blood collection, no history of tumor;

[0053] 3. Healthy female controls matched to the cases in age

[0054] The demographic and clinical data of these samples were systematically collected.

[0055] Example 2 Extraction and purification of peripheral blood DNA

[0056] Among the 25 eligible breast cancer patients and 10 healthy female controls, the two groups were balanced and comparable in age.

[0057] The specific steps are:

[0058] 1. Add hemolysis reagent (i.e., lysis buffer, 40 portions prepared as follows: 219.72 g sucrose, 2.02 g magnesium chloride, and 20 mL of Triton X-100 (amresco0694), then dilute to 2000 mL with TrisHCl solution, the same below) to the peripheral blood stored in a 2 mL cryovial, invert and mix thoroughly, and then transfer completely.

[0059] 2. Remove red blood cells: Fill the 5 mL centrifuge tube to 4 mL with hemolysis reagent, invert to mix, centrifuge at 4000 rpm for 10 minutes, and discard the supernatant. Add 4 mL of hemolysis reagent to the pellet, invert again to mix, and centrifuge at 4000 rpm for 10 minutes. Discard the supernatant.

[0060] 3. Extract DNA: Add 1 mL of extraction solution (each 300 mL contains 122.5 mL of 0.2 M sodium chloride, 14.4 mL of 0.5 M ethylenediaminetetraacetic acid, 15 mL of 10% sodium lauryl sulfate, and 148.1 mL of double-distilled water, the same below) and 8 μL of proteinase K to the precipitate, shake thoroughly on a shaker to mix, and incubate in a 37°C water bath overnight.

[0061] 4. Protein removal: Add 1 mL of saturated phenol and mix thoroughly (shake gently by hand for 15 minutes). Centrifuge at 4000 rpm for 10 minutes. Transfer the supernatant to a fresh 5 mL centrifuge tube. Add an equal volume of a mixture of chloroform and isoamyl alcohol (chloroform:isoamyl alcohol = 24:1, v / v, the same below) to the supernatant. Mix thoroughly (shake gently by hand for 15 minutes). Centrifuge at 4000 rpm for 10 minutes. Remove the supernatant and divide it into two 1.5 mL centrifuge tubes.

[0062] 5. DNA precipitation: Add 60 μL of 3M sodium acetate to the supernatant, then add an equal volume of ice-cold anhydrous ethanol to the supernatant. Shake gently up and down until a white flocculent precipitate is visible. Centrifuge at 12,000 rpm for 10 minutes.

[0063] 6. DNA washing: Add 1 mL of ice-cold anhydrous ethanol to the precipitate, centrifuge at 12,000 rpm for 10 min, discard the supernatant and vacuum dry or evaporate to dryness in a clean, dry environment.

[0064] 7. Measure the concentration: Usually 20-50 ng / μL DNA can be obtained, with a purity (UV 2600D: 2800D) of 1.8-2.0.

[0065] Example 3 Whole-exome detection of SNPs

[0066] The two groups of people in Example 2 were tested by whole exon chip to obtain relevant results.

[0067] 1. Library construction

[0068] Beijing Novogene Technology Co., Ltd. uses an Agilent liquid-phase microarray capture system to efficiently enrich human exome-wide DNA, followed by high-throughput, deep sequencing on the Illumina HiSeq platform. Library construction and capture experiments were performed using the Agilent SureSelect Human All Exon V5 kit, strictly following the recommended reagents and consumables in the manufacturer's instructions and following the latest optimized protocols.

[0069] Basic experimental procedures: Genomic DNA is randomly fragmented into 180-280bp fragments using a Covaris fragmentor. After end-repair and A-tailing, adapters are ligated to the fragment ends to prepare DNA libraries. After pooling, the libraries, which carry specific indices, are hybridized with up to 543,872 biotinylated probes in liquid phase. 334,378 exons from 20,965 genes are captured using streptavidin-containing magnetic beads. After PCR linear amplification, the libraries are quality-checked and ready for sequencing.

[0070] 2. Warehouse inspection

[0071] After the library construction is completed, Qubit2.0 is used for preliminary quantification, and the library is diluted to 1 ng / μL. Then, the insert size of the library is detected using Agilent 2100. Once the insert size meets the expectation, the effective concentration of the library is accurately quantified using Q-PCR (library effective concentration > 2 nM) to ensure the quality of the library.

[0072] 3. Sequencing

[0073] If the library is qualified, Illumina Hiseq platform sequencing will be performed according to the effective concentration of the library and data output requirements.

[0074] 4. Data analysis and processing

[0075] After data screening, in-depth processing, and bioinformatics sequence alignment, 53 SNP sites with significant differences in genotype distribution frequency between the "breast cancer case" group and the "healthy female control" group were finally identified as preferred sensitive sites. Among them, two SNP mutations located in the TCHH gene, the impact values of these site mutations on proteins are shown in Table 1:

[0076] Table 1 Effects of SNP mutation sites on protein

[0077] serial number ljb23_sift ljb23_pp2hvar ljb23_pp2hdiv ljb23_mt Susceptible SNP loci (1) 0.26,0.74,T 0.519,P 0.924,P 1,0.0,N Susceptible SNP loci (2) 0.81,0.19,T 0.766,P 0.991,D 1,0.0,N

[0078] After bioinformatics analysis, this site can be confirmed as a candidate marker for breast cancer.

[0079] Example 4 Further analysis of SNPs and breast cancer risk using risk scoring method

[0080] The inventors compared the genotype distribution frequencies of two sample groups (breast cancer cases and healthy women controls) and selected positively associated SNPs. Using the regression coefficients of individual SNPs in the whole-exome scan samples as weights, they further calculated risk scores and plotted receiver operating characteristic (ROC) curves to assess the sensitivity and specificity of the diagnosis, thereby determining the ability of these SNPs to predict breast cancer. A combined analysis of all SNP markers revealed that both sensitivity and specificity for the two susceptible SNPs exceeded 60%.

[0081] Therefore, the present inventors demonstrated that this locus marker can well distinguish healthy female controls from breast cancer patients.

[0082] Example 5 Genotyping of a single SNP

[0083] 1. Take DNA samples from 5 breast cancer patients and 5 healthy female controls as in Example 2;

[0084] 2. PCR amplification

[0085] The online primer design software provided by NCBI website https: / / www.ncbi.nlm.nih.gov / tools / primer-blast / index.cgi?LINK_LOC=BlastHomeAd was used to design single SNP-specific amplification primers for EME1:NM_001166131:exon9:c.G1738C as shown in Table 2.

[0086] Table 2 Primer sequences

[0087]

[0088] The PCR reaction system is shown in Table 3. The PCR amplification program was as follows: 95°C pre-denaturation for 10 min, 94°C denaturation for 15 s, 60°C annealing for 15 s, and 72°C extension for 30 s for 30 cycles, followed by a final extension at 72°C for 30 min. The cells were stored at 4°C and frozen at -20°C overnight.

[0089] Table 3 Reaction system

[0090] Components Addition amount 2×mix 25 μL Upstream primer (10uM) 3.0μL Downstream primer (10uM) 3.0μL template 5μL Add sterile distilled water to 50 μL

[0091] 3. Sequencing

[0092] After PCR amplification, 5 μL of the amplified product was subjected to electrophoresis on a 1% agarose gel for 30 minutes and stained for 20 minutes. The gel was then placed on a gel imager for observation. The correct amplified fragment was initially determined by comparing the fragment size with the marker. Amplified products that met the requirements were then purified using the Mag-Bind Oligonucleotide Purification Kit and performed according to the kit's instructions. Sequencing was performed using the ABI BigDye 3.1 Sequencing Kit and performed according to the kit's instructions on an ABI 3730 sequencer.

[0093] 4. Results Analysis

[0094] Using Chromas sequence analysis software, sequencing results were compared with standard sequences to identify SNP sites. By analyzing the base types at the SNP sites, the genotypes of the SNP sites were determined. The results showed that the variants at these two sites were real, further confirming that these two susceptibility SNPs can be used as auxiliary diagnostics for breast cancer detection, treatment, diagnosis, and prognosis assessment.

[0095] Example 6 Preparation of a Breast Cancer SNP Kit

[0096] Based on the primer set obtained in Example 5, a kit for breast cancer according to the present invention was assembled. The kit includes primer pairs for specifically amplifying two susceptible SNP sites, as shown in SEQ ID NO: 1 and SEQ ID NO: 2. The kit may also include common reagents required for corresponding PCR techniques, such as dNTPs, MgCl2, double-distilled water, Taq enzyme, etc. These common reagents are well known to those skilled in the art. In addition, standards and controls (such as standards for determining genotypes and blank controls) may also be included. The value of this kit lies in that it only requires peripheral blood and no other tissue samples. It detects SNPs using the most streamlined and specific primer pairs, and then assists in the diagnosis of breast cancer using the SNP spectrum. This kit is not only stable, convenient, and accurate, but also greatly improves the sensitivity and specificity of disease diagnosis. Therefore, putting this kit into practice can help guide diagnosis and more effective individualized treatment.

[0097] Example 7 Application of the kit

[0098] Between January 2010 and December 2015, blood samples were collected from 10 breast cancer patients before and after treatment at Shenzhen Second People's Hospital. The kit described in Example 6 was used to detect the genotypes of two susceptible SNPs in the TCHH gene to investigate the effectiveness of the kit for breast cancer detection. The results are as follows:

[0099] As shown in Table 4, the results of the kit for breast cancer detection were consistent with the clinical evaluation results by 90%.

[0100] Table 4 SNP site detection for pancreatic cancer efficacy evaluation

[0101] Patient number Susceptible SNP loci (1) Susceptible SNP loci (2) Clinical diagnosis results 1 C A Diagnosed with breast cancer 2 C A Diagnosed with breast cancer 3 C A Diagnosed with breast cancer 4 C A Diagnosed with breast cancer 5 C A Diagnosed with breast cancer 6 C G Undiagnosed breast cancer 7 C A Diagnosed with breast cancer 8 G G Undiagnosed breast cancer 9 C A Diagnosed with breast cancer 10 C A Diagnosed with breast cancer

[0102] Although the present invention has been described in detail above using general descriptions and specific embodiments, it will be apparent to those skilled in the art that modifications and improvements may be made based on the present invention. Therefore, such modifications and improvements, which do not depart from the spirit of the present invention, are intended to be within the scope of protection claimed herein. Sequence Listing <110> Shenzhen Second People's Hospital <120> A breast cancer detection kit <130> P16rxa98 <160> 2 <170> PatentIn version 3.5 <210> 1 <211> 18 <212> DNA <213> Artificial sequence <400> 1 tgcgcagtca ggaaccag 18 <210> 2 <211> 16 <212> DNA <213> Artificial sequence <400> 2 gttggccctc ctggcg 16

Claims

1. Use of a reagent comprising a primer pair for amplifying the genotypes of a TCHH gene susceptible SNP site (1) and a susceptible SNP site (2) in preparing a breast cancer detection product, wherein the specific information of the susceptible SNP site (1) is rs181128140, NM_007113: exon3: c.G4646C: p.R1549P, and the specific information of the susceptible SNP site (2) is NM_007113: exon3: c.G4678A: p.E1560K; The primer pair for amplifying the susceptible SNP site (1) and the susceptible SNP site (2) is shown as SEQ ID NO: 1 and SEQ ID NO:

2.

2. The use according to claim 1, characterized in that The breast cancer detection product is a kit.

3. The use according to claim 2, characterized in that The kit comprises reagents for detecting the genotype of a susceptible SNP site (1) and / or a susceptible SNP site (2) in a sample by sequencing.

4. The use according to claim 2, characterized in that The kit comprises reagents for detecting the genotype of a susceptible SNP site (1) and / or a susceptible SNP site (2) in a sample using a Taqman probe SNP detection method.

5. The use according to claim 2, characterized in that The kit is a kit for detecting the genotype of a susceptible SNP site (1) and / or a susceptible SNP site (2) in a sample using a PCR-single-strand conformation polymorphism method.

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