CN108676890B - Female breast malignant tumor susceptibility prediction kit and system - Google Patents

Abstract

The present invention relates to a kit for predicting the susceptibility of female breast cancer, comprising the following components: STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer, STR-6 primer Further, it can also include: PCR amplification reaction solution, LIZ-500 molecular weight internal standard, deionized formamide. The female breast cancer susceptibility prediction kit of the present invention can be used for diagnosis and susceptibility prediction of female breast cancer. The present invention also provides a system for predicting the susceptibility of female breast cancer.

Description

A kit and system for predicting the susceptibility of female breast cancer

technical field

The present invention relates to the field of biomedicine. Specifically, it relates to a female breast cancer susceptibility prediction kit and a female breast cancer susceptibility prediction system. More specifically, the present invention relates to a test kit for detecting the STR of the susceptibility-related gene of female breast cancer susceptibility by the method of short tandem repeats (Short tandem repeats, referred to as STR) site fragment analysis, and by combining the discriminant analysis statistical method, In this way, an early warning of the susceptibility of female breast cancer in the subject is carried out.

Background technique

Tumor is a disease closely related to genetics. To study the molecular genetic basis of tumor, and then propose tumor-specific genetic markers, it is hoped that it can be used for routine detection, clinical diagnosis, personalized treatment, and follow-up. etc. to provide a simple and feasible method. However, the individual differences of patients and the intersection of related biomolecular events at different developmental stages have brought great difficulties to this work.

Female breast cancer is mainly from breast duct and lobular epithelium, accounting for more than 95% of female breast cancer; followed by soft tissue sarcoma from breast mesenchymal tissue, less than 5%. The occurrence of this type of disease is related to endocrine factors, diet and obesity, radiation exposure, milk factors and other factors. Therefore, predicting the susceptibility of breast cancer in the examined population is helpful to improve the awareness of disease risk, reduce the pathogenesis by adjusting diet and working environment, and at the same time help patients to detect and treat early.

A large number of studies have shown that genetic polymorphisms of tumor-related genes play a key role in the occurrence and development of malignant tumors. However, the occurrence and development of tumors is a very complex process, and it is obviously impossible and unscientific to use the changes of a single molecular genetic marker to diagnose the disease. With the existing technical means, it is still not possible to carry out a relatively accurate early warning of tumor susceptibility only through genetic information, and the current early identification and prediction methods for tumors still need to be improved. The invention relates to a kit for early warning of the susceptibility of female breast cancer by combining the STR site fragment analysis method to detect a plurality of STR sites with high correlation with the occurrence of female breast cancer, combined with the discriminant analysis statistical method.

SUMMARY OF THE INVENTION

In order to solve the above-mentioned problems in the prior art, the present invention relates to the joint detection of multiple STR loci that are highly correlated with the occurrence of female breast cancer through the STR locus fragment analysis method, combined with the discriminant analysis statistical method, and the detection of female breast cancer. Early warning of malignancy susceptibility.

The present invention is accomplished based on the inventors' following findings: the inventors analyze the STR of the genomic DNA of the female breast malignant tumor subjects and the healthy control subjects, and verify the results in a large number of female breast malignant tumor samples and control samples , found that the number of short tandem sequence repeats of each independent STR locus was not significantly correlated with the female breast cancer in the subject, while the combination of the number of short tandem sequence repeats of some specific STR loci was associated with the female breast cancer in the subject. Malignant tumors are closely related.

To this end, the present invention proposes a set of isolated STR loci that are highly associated with female breast malignancies. According to embodiments of the present invention, these isolated STR loci comprise the nucleotide sequences shown in STR-1 to STR-6 (Table 1). Using these isolated STR loci as a reference, the susceptibility of female breast cancer can be effectively predicted.

Table 1

Site code starting point gene short tandem sequence STR-1 X chromosome, position 66657655 AR CAG STR-2 Chromosome 4, position 55633758 Bat-25 T STR-3 Chromosome 5, position 111646983 D5S346 GT STR-4 Chromosome 6, position 151806531 ER1 TA STR-5 Chromosome 14, position 64253561 ER2 TG STR-6 Chromosome 4, position 154587748 FGA AAAG

For the detailed description of the above-mentioned STR sites, those skilled in the art can log in to relevant databases (such as GeneBank, Nucleotide, etc.) to obtain, and will not be repeated here. The inventors have surprisingly found that by analyzing the cell genome of the subject to obtain the number of repetitions of the short tandem sequence of each STR site above, and using the number of repetitions as an independent variable to perform statistical analysis methods such as discriminant analysis, it is possible to detect malignant breast cancer in women. Early warning of tumor susceptibility.

On this basis, one of the technical problems solved by the present invention is to provide a female breast cancer susceptibility prediction kit, which includes the following components: STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primers, STR-5 primers, and STR-6 primers are used to amplify the target fragments containing the short tandem sequences listed in Table 1, respectively, to determine the number of repetitions of the short tandem sequences.

Preferably, the female breast cancer susceptibility prediction kit of the present invention further comprises: PCR amplification reaction solution, LIZ-500 molecular weight internal standard, and deionized formamide.

In the female breast cancer susceptibility prediction kit of the present invention, preferably, the sequences of the above-mentioned STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer and STR-6 primer are as follows As shown in Table 2, more preferably, the concentration of each of the above primers is 10 μM:

Table 2

In Table 2, HEX, FAM, and ROX are all fluorescent groups for labeling the 5' end, HEX is hexachloro-6-methylfluorescein, FAM is 6-carboxyfluorescein, and ROX is ROX reference dye.

In the female breast cancer susceptibility prediction kit of the present invention, preferably, the PCR amplification reaction solution is a mixture of the following reagents: TaqDNA polymerase (5U/μL), Tris-HCl (100mM, at 25°C) pH 8.8), KCl (500 mM), ethylphenyl polyethylene glycol (0.8% (v/v)), _MgCl2 (25 mM), dNTPs (10 mM), deionized water.

More preferably, the PCR amplification reaction solution is stored at -20°C.

In the female breast cancer susceptibility prediction kit of the present invention, preferably, the LIZ-500 molecular weight internal standard can be stored at -20°C;

In the female breast cancer susceptibility prediction kit of the present invention, preferably, the deionized formamide can be stored at 2-8°C.

Preferably, the female breast cancer susceptibility prediction kit of the present invention further includes instructions for use.

The instructions for use describe a method for using the above-mentioned kit for predicting the susceptibility of female breast cancer, which includes the following steps:

(1) Extract sample DNA;

(2) PCR reaction

(2-1) Take out the STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer, STR-6 primer, and PCR amplification reaction solution from the refrigerator, and equilibrate to room temperature. After the components are fully dissolved, centrifuge quickly for 10 seconds;

(2-2) Take 30-300ng of sample DNA, add 60μL of PCR amplification reaction solution, add deionized water to make up to 115.2μL, mix well, centrifuge quickly for 10 seconds, and then dispense the mixture at 19.2μL/well to 6 PCR reaction tubes;

(2-3) Add STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer, and STR-6 primer to step (2-2) at 0.8 μL/well respectively. 6 PCR reaction tubes; cover the PCR reaction tube caps, record the sample loading situation, centrifuge quickly for 10 seconds, then transfer the PCR reaction tubes to the corresponding position of the sample tank of the PCR amplifier, record the placement sequence, and start PCR amplification Amplification reaction conditions: 95°C for 3 minutes; 95°C for 30 seconds, 60°C for 30 seconds, 72°C for 30 seconds, 10 cycles; 95°C for 30 seconds, 55°C for 30 seconds, 72°C for 30 seconds, 20 cycles ; 6 minutes at 72°C to obtain 6 sets of PCR amplification products;

(3) STR fragment analysis

(3-1) Take 990 μL of deionized formamide, add 10 μL of LIZ-500 molecular weight internal standard, mix well, centrifuge quickly for 10 seconds, add 10 μL/well to sequencing reaction tubes, and centrifuge quickly for 10 seconds;

(3-2) Add 1 μL/well of the above-mentioned 6 sets of PCR amplification products to 6 sequencing reaction tubes, and centrifuge quickly for 10 seconds; then transfer the sequencing reaction tubes to the corresponding position of the sample tank of the PCR amplifier, 98°C Heating for 5 minutes. Immediately after the end of the program, the sequencing reaction tube was placed on the ice-water mixture and rapidly cooled to 0 °C and centrifuged rapidly for 10 seconds; then the sequencing reaction tube was transferred to the corresponding position of the sample tank of the STR locus fragment analyzer, and recorded and placed sequence, perform fragment analysis and detection;

(4) Analysis and judgment of results

(4-1) According to the fragment analysis results, record the fragment lengths of the two alleles of STR-1, STR-2, STR-3, STR-4, STR-5, and STR-6 respectively:

The smaller fragment length value of the two alleles of STR-1 is recorded as L ₁ , and the larger fragment length value of the two alleles of STR-1 is recorded as L ₂ ;

The smaller fragment length value of the two alleles of STR-2 is recorded as L ₃ , and the larger fragment length value of the two alleles of STR-2 is recorded as L ₄ ;

The smaller fragment length value of the two alleles of STR-3 is recorded as L ₅ , and the larger fragment length value of the two alleles of STR-3 is recorded as L ₆ ;

The smaller fragment length value of the two alleles of STR-4 is recorded as L ₇ , and the larger fragment length value of the two alleles of STR-4 is recorded as L ₈ ;

The smaller fragment length value of the two alleles of STR-5 is recorded as L ₉ , and the larger fragment length value of the two alleles of STR-5 is recorded as L ₁₀ ;

The smaller fragment length value of the two alleles of STR-6 is recorded as L ₁₁ , and the larger fragment length value of the two alleles of STR-6 is recorded as L ₁₂ ;

(4-2) The number of repetitions of the short tandem sequence is calculated according to the fragment length and the following formula, and denoted as X ₁ -X ₁₂ , where round represents rounding to an integer:

X ₁ =round[(L ₁ -191)/3]; X ₂ =round[(L ₂ -191)/3];

X ₃ =round(L ₃ -379); X ₄ =round(L ₄ -379);

X ₅ =round[(L ₅ -202)/2]; X ₆ =round[(L ₆ -202)/2];

X ₇ =round[(L ₇ -359)/2]; X ₈ =round[(L ₈ -359)/2];

X ₉ =round[(L ₉ -278)/2]; X ₁₀ =round[(L ₁₀ -278)/2];

X ₁₁ =round[(L ₁₁ -200)/4]; X ₁₂ =round[(L ₁₂ -200)/4];

(4-3) Substitute the repetition times of the above short series sequence into the preset discriminant function:

F _BC = 12.108X ₁ -0.933X ₂ +15.490X ₃ +112.846X ₄ +2.606X ₅ +0.178X ₆ +6.318X ₇ -1.147X ₈ +2.405X ₉ -4.504X ₁₀ +2.549X ₁₁ +5.507X ₁₂ -1783.160

F _BN = 12.504X ₁ -1.212X ₂ +15.869X ₃ +115.560X ₄ +2.561X ₅ -0.026X ₆ +6.887X ₇ -1.615X ₈ +2.619X ₉ -4.518X ₁₀ +2.486X ₁₁ +5.478X ₁₂ -1860.413

(4-4) Prediction of susceptibility to female breast cancer:

Comparing the F _BC value and the F _BN value, if F _BC > F _BN , it is predicted that the probability of the subject suffering from female breast cancer is ≥90.0%; if F _BC ≤ F _BN , it is predicted that the subject will not suffer from female breast malignant tumor The odds of ≥77.8%.

In the present invention,

Preferably, the extraction of sample DNA in step (1) can use a purchased commercial genomic DNA extraction kit and operate according to the kit instructions. The sample may be whole blood of the subject.

Preferably, the rotational speed of all centrifugation in the method of use is preferably 3000 g/min.

The probability of suffering from female breast cancer in the present invention is the sum of the probability of having suffered from female breast cancer and the probability of suffering from female breast cancer in the future. Therefore, it can be used not only for the diagnosis of female breast cancer, but also for the risk warning of female breast cancer in the future. , reduce the risk of female breast cancer.

The second technical problem solved by the present invention is to provide a method for predicting the susceptibility of female breast cancer, that is, using the above-mentioned kit and operating according to the method described in the specification.

The third technical problem solved by the present invention is to provide the application of the female breast cancer susceptibility prediction kit in the preparation of a diagnosis product for female breast cancer.

The fourth technical problem solved by the present invention is to provide a female breast cancer susceptibility prediction system, which includes:

A device for obtaining the number of repetitions of the short tandem sequence of the STR site of the sample DNA;

A data processing and determination device, which includes the following modules:

The data input module is used to input the age, gender, and repetition times of the short tandem sequence of the STR locus;

The database management module is used for the operation management of data storage, modification, deletion, query and printing;

The data calculation module is used to calculate the discriminant function result according to the repetition times of the short tandem sequence of the STR site in the data input module;

The analysis discriminant and result output module is used to compare the discriminant function results, so as to predict the susceptibility of female breast cancer, and output the result.

in,

The number of repetitions of the short tandem sequences at the STR site is the number of repetitions of the following 6 pairs of short tandem sequences at the STR site:

Site code starting point gene short tandem sequence STR-1 X chromosome, position 66657655 AR CAG STR-2 Chromosome 4, position 55633758 Bat-25 T STR-3 Chromosome 5, position 111646983 D5S346 GT STR-4 Chromosome 6, position 151806531 ER1 TA STR-5 Chromosome 14, position 64253561 ER2 TG STR-6 Chromosome 4, position 154587748 FGA AAAG

The discriminant function includes:

First discriminant function F _BC =12.108X ₁ -0.933X ₂ +15.490X ₃ +112.846X ₄ +2.606X ₅ +0.178X ₆ +6.318X ₇ -1.147X ₈ +2.405X ₉ -4.504X ₁₀ +2.549X ₁₁ +5.507X ₁₂ -1783.160

Second discriminant function F _BN =12.504X ₁ -1.212X ₂ +15.869X ₃ +115.560X ₄ +2.561X ₅ -0.026X ₆ +6.887X ₇ -1.615X ₈ +2.619X ₉ -4.518X ₁₀ +2.486X ₁₁ +5.478X ₁₂ -1860.413

In the discriminant function,

X ₁ is the number of repetitions of the short tandem sequence of the smaller fragment in the two alleles of STR-1;

X ₂ is the number of repetitions of the short tandem sequence of the larger fragment in the two alleles of STR-1;

X ₃ is the number of repetitions of the short tandem sequence of the smaller fragment in the two alleles of STR-2;

X ₄ is the number of repetitions of the short tandem sequence of the larger fragment in the two alleles of STR-2;

X ₅ is the number of repetitions of the short tandem sequence of the smaller fragment in the two alleles of STR-3;

X ₆ is the number of repetitions of the short tandem sequence of the larger fragment in the two alleles of STR-3;

X ₇ is the number of repetitions of the short tandem sequence of the smaller fragment in the two alleles of STR-4;

X ₈ is the number of repetitions of the short tandem sequence of the larger fragment in the two alleles of STR-4;

X ₉ is the number of repetitions of the short tandem sequence of the smaller fragment in the two alleles of STR-5;

X ₁₀ is the number of repetitions of the short tandem sequence of the larger fragment in the two alleles of STR-5;

X ₁₁ is the number of repetitions of the short tandem sequence of the smaller fragment in the two alleles of STR-6;

X ₁₂ is the number of repetitions of the short tandem sequence of the larger fragment in the two alleles of STR-6;

Among them, X ₁ ^- X ₁₂ are calculated according to the fragment length and the following formula, where round represents rounding to an integer:

X ₁ =round[(L ₁ -191)/3]; X ₂ =round[(L ₂ -191)/3];

X ₃ =round(L ₃ -379); X ₄ =round(L ₄ -379);

X ₅ =round[(L ₅ -202)/2]; X ₆ =round[(L ₆ -202)/2];

X ₇ =round[(L ₇ -359)/2]; X ₈ =round[(L ₈ -359)/2];

X ₉ =round[(L ₉ -278)/2]; X ₁₀ =round[(L ₁₀ -278)/2];

X ₁₁ =round[(L ₁₁ -200)/4]; X ₁₂ =round[(L ₁₂ -200)/4];

In X ₁ ^- X ₁₂ , L ₁ is the smaller fragment length value of the two alleles of STR-1, and L ₂ is the larger fragment length value of the two alleles of STR-1;

L ₃ is the smaller fragment length value of the two alleles of STR-2, and L ₄ is the larger fragment length value of the two alleles of STR-2;

L ₅ is the smaller fragment length value of the two alleles of STR-3, and L ₆ is the larger fragment length value of the two alleles of STR-3;

L ₇ is the smaller fragment length value of the two alleles of STR-4, and L ₈ is the larger fragment length value of the two alleles of STR-4;

L ₉ is the smaller fragment length value of the two alleles of STR-5, and L ₁₀ is the larger fragment length value of the two alleles of STR-5;

L ₁₁ is the smaller fragment length value of the two alleles of STR-6, and L ₁₂ is the larger fragment length value of the two alleles of STR-6;

The analysis, discrimination and result output module compares the calculation result of the first discriminant function F _BC with the calculation result of the second discriminant function F _BN , and if F _BC > _{FB BN} , it outputs “the subject suffers from a female breast cancer. If F _BC ≤ F _BN , output the prediction result of "the probability of the subject not suffering from female breast cancer ≥ 77.8%".

The device for obtaining the repetition times of the short tandem sequence of the STR site of the sample DNA may include a STR site fragment analyzer, a PCR amplifier, etc.; the data processing and determination device may be a computer or other equipment.

The fifth technical problem solved by the present invention is to provide the application of the female breast cancer susceptibility prediction system in the preparation of female breast cancer prediction products, female breast cancer diagnosis products, and breast health auxiliary products.

The sixth technical problem solved by the present invention is to provide a product for predicting female breast cancer, a product for diagnosing female breast cancer, or a breast health auxiliary product, which includes the system for predicting the susceptibility of female breast cancer .

The test material used in the present invention is human genomic DNA. In theory, human genomic DNA will not change in a person's life. Human genomic DNA encodes all life activities of human beings, so theoretically, by detecting genomic DNA, the risk of a subject suffering from a certain disease can be predicted early, even at birth. The present invention is based on this theory, so using The kit and system of the present invention can not only play the role of early warning, but also play the role of diagnosis.

The occurrence and development of tumors is a very complex process. To study the molecular genetic basis of tumors, it is hoped that it can provide simple and feasible methods for routine detection, clinical diagnosis, personalized treatment, and follow-up of the disease after recovery. However, the individual differences of patients and the intersection of related biomolecular events at different developmental stages have brought great difficulties to this work. Using a single molecular genetic change to diagnose a tumor is clearly impossible and unscientific. The inventors applied modern molecular biology technology to jointly analyze multiple STRs in the genomic DNA of the subject, combined with statistical analysis methods such as discriminant analysis, thereby inventing a kit for early warning of female breast cancer susceptibility.

Description of drawings

FIG. 1 is a schematic diagram of the modules included in the data processing and determination device in the system for predicting the susceptibility to malignant tumors of the female breast according to the present invention.

Detailed ways

The specific embodiments of the present invention will be described below with reference to the accompanying drawings and examples, so as to better understand the present invention. The experimental methods used in the following examples are conventional methods unless otherwise specified. The following describes in detail the embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein the same or similar reference numerals refer to the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary, only used to explain the present invention, and should not be construed as a limitation of the present invention.

The PCR amplification instrument in the embodiment is the Mastercycler nexus amplification instrument (purchased from Eppendorf, USA);

The STR locus fragment analyzer in the examples is a 3730XL sequencing analyzer (purchased from ABI, USA);

The DNA extraction kit in the embodiment is blood DNAout kit (purchased from Beijing Tianenze Company);

The rotational speed of all centrifugation in the examples was 3000 g/min.

Example 1 A system for predicting the susceptibility of female breast cancer

A female breast cancer susceptibility prediction system, comprising:

A device for obtaining the number of repetitions of the short tandem sequence of the STR site of the sample DNA;

A data processing and determination device, which includes the following modules (as shown in Figure 1):

The data input module is used to input the age, gender, and repetition times of the short tandem sequence of the STR locus;

The database management module is used for the operation management of data storage, modification, deletion, query and printing;

The data calculation module is used to calculate the discriminant function result according to the repetition times of the short tandem sequence of the STR site in the data input module;

The analysis discriminant and result output module is used to compare the discriminant function results, so as to predict the susceptibility of female breast cancer, and output the result.

in,

The number of repetitions of the short tandem sequences at the STR site is the number of repetitions of the following 6 pairs of short tandem sequences at the STR site:

The discriminant function includes:

First discriminant function F _BC =12.108X ₁ -0.933X ₂ +15.490X ₃ +112.846X ₄ +2.606X ₅ +0.178X ₆ +6.318X ₇ -1.147X ₈ +2.405X ₉ -4.504X ₁₀ +2.549X ₁₁ +5.507X ₁₂ -1783.160

Second discriminant function F _BN =12.504X ₁ -1.212X ₂ +15.869X ₃ +115.560X ₄ +2.561X ₅ -0.026X ₆ +6.887X ₇ -1.615X ₈ +2.619X ₉ -4.518X ₁₀ +2.486X ₁₁ +5.478X ₁₂ -1860.413

In the discriminant function,

X ₁ is the number of repetitions of the short tandem sequence of the smaller fragment in the two alleles of STR-1;

X ₂ is the number of repetitions of the short tandem sequence of the larger fragment in the two alleles of STR-1;

X ₃ is the number of repetitions of the short tandem sequence of the smaller fragment in the two alleles of STR-2;

X ₄ is the number of repetitions of the short tandem sequence of the larger fragment in the two alleles of STR-2;

X ₅ is the number of repetitions of the short tandem sequence of the smaller fragment in the two alleles of STR-3;

X ₆ is the number of repetitions of the short tandem sequence of the larger fragment in the two alleles of STR-3;

X ₇ is the number of repetitions of the short tandem sequence of the smaller fragment in the two alleles of STR-4;

X ₈ is the number of repetitions of the short tandem sequence of the larger fragment in the two alleles of STR-4;

X ₉ is the number of repetitions of the short tandem sequence of the smaller fragment in the two alleles of STR-5;

X ₁₀ is the number of repetitions of the short tandem sequence of the larger fragment in the two alleles of STR-5;

X ₁₁ is the number of repetitions of the short tandem sequence of the smaller fragment in the two alleles of STR-6;

_X12 is the number of repetitions of the short tandem sequence of the larger fragment of the two alleles of STR-6.

Among them, X ₁ ^- X ₁₂ are calculated according to the fragment length and the following formula, where round represents rounding to an integer:

X ₁ =round[(L ₁ -191)/3]; X ₂ =round[(L ₂ -191)/3];

X ₃ =round(L ₃ -379); X ₄ =round(L ₄ -379);

X ₅ =round[(L ₅ -202)/2]; X ₆ =round[(L ₆ -202)/2];

X ₇ =round[(L ₇ -359)/2]; X ₈ =round[(L ₈ -359)/2];

X ₉ =round[(L ₉ -278)/2]; X ₁₀ =round[(L ₁₀ -278)/2];

X ₁₁ =round[(L ₁₁ -200)/4]; X ₁₂ =round[(L ₁₂ -200)/4];

In X ₁ ^- X ₁₂ , L ₁ is the smaller fragment length value of the two alleles of STR-1, and L ₂ is the larger fragment length value of the two alleles of STR-1;

L ₃ is the smaller fragment length value of the two alleles of STR-2, and L ₄ is the larger fragment length value of the two alleles of STR-2;

L ₅ is the smaller fragment length value of the two alleles of STR-3, and L ₆ is the larger fragment length value of the two alleles of STR-3;

L ₇ is the smaller fragment length value of the two alleles of STR-4, and L ₈ is the larger fragment length value of the two alleles of STR-4;

L ₉ is the smaller fragment length value of the two alleles of STR-5, and L ₁₀ is the larger fragment length value of the two alleles of STR-5;

L ₁₁ is the smaller fragment length value of the two alleles of STR-6, and L ₁₂ is the larger fragment length value of the two alleles of STR-6;

The analysis, discrimination and result output module compares the calculation result of the first discriminant function F _BC with the calculation result of the second discriminant function F _BN , and if F _BC > _{FB BN} , it outputs “the subject suffers from a female breast cancer. If F _BC ≤ F _BN , output the prediction result of "the probability of the subject not suffering from female breast cancer ≥ 77.8%".

Example 2 A kit for predicting the susceptibility of female breast cancer

A kit for predicting the susceptibility of female breast cancer, comprising the following components: STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer, STR-6 primer, PCR amplification Addition of reaction solution, LIZ-500 molecular weight internal standard, deionized formamide, instruction manual.

The concentrations of the above STR-1 primers, STR-2 primers, STR-3 primers, STR-4 primers, STR-5 primers, and STR-6 primers are all 10 μM, and the primer sequences are shown in the following table:

The PCR amplification reaction solution was a mixture of the following reagents: TaqDNA polymerase (5U/μL), Tris-HCl (100mM, pH 8.8 at 25°C), KCl (500mM), ethylphenyl polyethylene glycol (0.8 % (v/v)), _MgCl2 (25 mM), dNTPs (10 mM), deionized water.

The PCR amplification reaction solution was stored at -20°C; the LIZ-500 molecular weight internal standard was stored at -20°C; and the deionized formamide was stored at 2-8°C.

The above-mentioned kit also includes instructions for use.

Example 3 Using the system of Example 1 and the kit of Example 2 to predict the risk of female breast cancer in the subject

Subject: female, 52 years old, visiting the Department of Breast Surgery of the Second Hospital of Jilin University, fully informed of the purpose and use of the examination, signed an informed consent form, and collected 1 mL of anticoagulant through a peripheral vein.

Using the kit of Example 2, the following steps were performed according to the method described in the kit instructions:

(1) Extract DNA from the sample: extract genomic DNA from blood using a DNA extraction kit;

(2) PCR reaction

(2-1) Take out the STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer, STR-6 primer, and PCR amplification reaction solution from the refrigerator, and equilibrate to room temperature. After the components are fully dissolved, centrifuge quickly for 10 seconds;

(2-2) Take 100ng of sample DNA, add 60μL of PCR amplification reaction solution, add deionized water to make up to 115.2μL, mix well, centrifuge quickly for 10 seconds, and then divide the mixture into 6 cells at 19.2μL/well PCR reaction tube;

(2-3) Add STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer, and STR-6 primer to step (2-2) at 0.8 μL/well respectively. 6 PCR reaction tubes; cover the PCR reaction tube caps, record the sample loading situation, centrifuge quickly for 10 seconds, then transfer the PCR reaction tubes to the corresponding position of the sample tank of the PCR amplifier, record the placement sequence, and start PCR amplification Amplification reaction conditions: 95°C for 3 minutes; 95°C for 30 seconds, 60°C for 30 seconds, 72°C for 30 seconds, 10 cycles; 95°C for 30 seconds, 55°C for 30 seconds, 72°C for 30 seconds, 20 cycles ; 6 minutes at 72°C to obtain 6 sets of PCR amplification products;

(3) STR fragment analysis

(3-1) Take 990 μL of deionized formamide, add 10 μL of LIZ-500 molecular weight internal standard, mix well, centrifuge quickly for 10 seconds, add 10 μL/well to sequencing reaction tubes, and centrifuge quickly for 10 seconds;

(3-2) Add 1 μL/well of the above-mentioned 6 sets of PCR amplification products to 6 sequencing reaction tubes, and centrifuge quickly for 10 seconds; then transfer the sequencing reaction tubes to the corresponding position of the sample tank of the PCR amplifier, 98°C Heating for 5 minutes. Immediately after the end of the program, the sequencing reaction tube was placed on the ice-water mixture and rapidly cooled to 0 °C and centrifuged rapidly for 10 seconds; then the sequencing reaction tube was transferred to the corresponding position of the sample tank of the STR locus fragment analyzer, and recorded and placed sequence, perform fragment analysis and detection;

(4) Analysis and judgment of results

(4-1) According to the fragment analysis results, record the fragment lengths of the two alleles of STR-1, STR-2, STR-3, STR-4, STR-5, and STR-6 respectively: STR-1 two The smaller fragment length value of the two alleles is recorded as L ₁ , the larger fragment length value of the two alleles of STR-1 is recorded as L ₂ ; the smaller fragment length of the two alleles of STR-2 is recorded as L 2 . The value is recorded as L ₃ , the larger fragment length value of the two alleles of STR-2 is recorded as L ₄ ; the smaller fragment length value of the two alleles of STR-3 is recorded as L ₅ , and the two STR-3 alleles are recorded as L 5 . The larger fragment length value of the two alleles is recorded as L ₆ ; the smaller fragment length value of the two alleles of STR-4 is recorded as L ₇ , and the larger fragment length of the two alleles of STR-4 is recorded as L 7 . The value is recorded as L ₈ ; the smaller fragment length value of the two alleles of STR-5 is recorded as L ₉ , and the larger fragment length value of the two alleles of STR-5 is recorded as L ₁₀ ; The smaller fragment length value among the two alleles was recorded as L ₁₁ , and the larger fragment length value among the two alleles of STR-6 was recorded as L ₁₂ ; the results showed: L ₁ =273.59, L ₂ =276.27, L 12 ₃ = _403.24 , L4= _403.24 , L5= _229.33 , L6= _246.78 , _L7 =384.99, L8=396.6, L9= _312.13 , L10= _320.45 , L11= _252.54 , L12= _264.19 .

(4-2) Calculated according to the length of the fragment and the following formula, denoted as X ₁ -X ₁₂ , where round represents rounding to an integer:

X ₁ =round[(L ₁ -191)/3]=28; X ₂ =round[(L ₂ -191)/3]=28;

X ₃ =round(L ₃ -379)=24; X ₄ =round(L ₄ -379)=24;

X ₅ =round[(L ₅ -202)/2]=14; X ₆ =round[(L ₆ -202)/2]=22;

X ₇ =round[(L ₇ -359)/2]=13; X ₈ =round[(L ₈ -359)/2]=19;

X ₉ =round[(L ₉ -278)/2]=17; X ₁₀ =round[(L ₁₀ -278)/2]=21;

X ₁₁ =round[(L ₁₁ -200)/4]=13; X ₁₂ =round[(L ₁₂ -200)/4]=16.

(4-3) Using the computer running the system for predicting the susceptibility of female breast malignant tumor described in Example 1, to predict the susceptibility of the subject to suffer from female breast malignant tumor:

The age, gender, and repetition times of the short tandem sequence of the STR site are input into the system through the data input module, and the result of the discriminant function is calculated through the data calculation module:

First discriminant function F _BC =12.108X ₁ -0.933X ₂ +15.490X ₃ +112.846X ₄ +2.606X ₅ +0.178X ₆ +6.318X ₇ -1.147X ₈ +2.405X ₉ -4.504X ₁₀ +2.549X ₁₁ +5.507X ₁₂ -1783.160＝1778.095

Second discriminant function F _BN =12.504X ₁ -1.212X ₂ +15.869X ₃ +115.560X ₄ +2.561X ₅ -0.026X ₆ +6.887X ₇ -1.615X ₈ +2.619X ₉ -4.518X ₁₀ +2.486X ₁₁ +5.478X ₁₂ -1860.413=1773.798

After the analysis, discrimination and result output module compares the F _BC value and the F _BN value, F _BC > F _BN , and outputs the prediction result of "the probability of the subject suffering from female breast cancer ≥ 90.0%".

The subject underwent modified radical mastectomy for breast cancer after seeing a doctor. The pathological examination was confirmed as invasive ductal carcinoma of the breast, and the clinical diagnosis results were consistent with the prediction results of the kit of the present invention.

Example 4 Using the system of Example 1 and the kit of Example 2 to predict the risk of female breast cancer in the subject

Subject: female, 60 years old, was admitted to the Department of Breast Surgery of the Second Hospital of Jilin University, underwent mammary duct endoscopy, was fully informed of the purpose and purpose of the examination, signed the informed consent on the premise of her own voluntariness, and passed the peripheral vein Collect 1 mL of anticoagulant.

Referring to the prediction method of Example 3, the same processing and testing were performed on the blood samples, and the results showed: L ₁ =265.48, L ₂ =270.86, L ₃ =404.17, L ₄ =404.17, L ₅ =228.62, L ₆ =228.62, L ₇ =398.52, L ₈ =404.64, L ₉ =324.6, L ₁₀ =330.99, L ₁₁ =255.66, L ₁₂ =263.75.

Calculated according to the fragment length and the following formula, denoted as X ₁ -X ₁₂ , where round represents rounding to an integer:

X ₁ =round[(L ₁ -191)/3]=25; X ₂ =round[(L ₂ -191)/3]=27;

X ₃ =round(L ₃ -379)=25; X ₄ =round(L ₄ -379)=25;

X ₅ =round[(L ₅ -202)/2]=13; X ₆ =round[(L ₆ -202)/2]=13;

X ₇ =round[(L ₇ -359)/2]=20; X ₈ =round[(L ₈ -359)/2]=23;

X ₉ =round[(L ₉ -278)/2]=23; X ₁₀ =round[(L ₁₀ -278)/2]=26;

X ₁₁ =round[(L ₁₁ -200)/4]=14; X ₁₂ =round[(L ₁₂ -200)/4]=16.

Using the computer running the system for predicting the susceptibility of female breast cancer described in Example 1, the susceptibility prediction of the subject to suffering from female breast cancer:

The age, gender, and repetition times of the short tandem sequence of the STR site are input into the system through the data input module, and the result of the discriminant function is calculated through the data calculation module:

First discriminant function F _BC =12.108X ₁ -0.933X ₂ +15.490X ₃ +112.846X ₄ +2.606X ₅ +0.178X ₆ +6.318X ₇ -1.147X ₈ +2.405X ₉ -4.504X ₁₀ +2.549X ₁₁ +5.507X ₁₂ -1783.160＝1900.929

The second discriminant function F _BN =12.504X ₁ -1.212X ₂ +15.869X ₃ +115.560X ₄ +2.561X ₅ -0.026X ₆ +6.887X ₇ -1.615X ₈ +2.619X ₉ -4.518X ₁₀ +2.486X ₁₁ +5.478X ₁₂ -1860.413＝1903.959

After the analysis, discrimination and result output module compares the F _BC value and the F _BN value, F _BC ≤ F _BN , and outputs the prediction result of "the probability of the subject not suffering from female breast cancer ≥ 77.8%".

The subject was diagnosed with intraductal papilloma after visiting a doctor, which is a benign tumor, and the clinical diagnosis result is consistent with the prediction result of the kit of the present invention.

Example 5 Using the system of Example 1 and the kit of Example 2 to predict the risk of female breast cancer in the subject

Subject: female, 59 years old, was admitted to the Department of Breast Surgery of the Second Hospital of Jilin University, and underwent modified radical mastectomy for breast cancer. She was fully informed of the purpose and use of the examination, signed an informed consent form on the premise of her own voluntariness, and was administered through a peripheral vein. Collect 1 mL of anticoagulant.

Referring to the prediction method of Example 3, the same processing and testing were performed on the blood samples, and the results showed: L ₁ =268.46, L ₂ =287.38, L ₃ =403.19, L ₄ =403.19, L ₅ =229.08, L ₆ =229.08, L ₇ =383.05, L ₈ =388.80, L ₉ =312.05, L ₁₀ =318.12, L ₁₁ =253.32, L ₁₂ =260.50.

Calculated according to the fragment length and the following formula, denoted as X ₁ -X ₁₂ , where round represents rounding to an integer:

X ₁ =round[(L ₁ -191)/3]=23; X ₂ =round[(L ₂ -191)/3]=28;

X ₃ =round(L ₃ -379)=24; X ₄ =round(L ₄ -379)=24;

X ₅ =round[(L ₅ -202)/2]=21; X ₆ =round[(L ₆ -202)/2]=22;

X ₇ =round[(L ₇ -359)/2]=12; X ₈ =round[(L ₈ -359)/2]=20;

X ₉ =round[(L ₉ -278)/2]=16; X ₁₀ =round[(L ₁₀ -278)/2]=17;

X ₁₁ =round[(L ₁₁ -200)/4]=13; X ₁₂ =round[(L ₁₂ -200)/4]=13;

Using the computer running the system for predicting the susceptibility of female breast cancer described in Example 1, the susceptibility prediction of the subject to suffering from female breast cancer:

The age, gender, and repetition times of the short tandem sequence of the STR site are input into the system through the data input module, and the result of the discriminant function is calculated through the data calculation module:

First discriminant function F _BC =12.108X ₁ -0.933X ₂ +15.490X ₃ +112.846X ₄ +2.606X ₅ +0.178X ₆ +6.318X ₇ -1.147X ₈ +2.405X ₉ -4.504X ₁₀ +2.549X ₁₁ +5.507X ₁₂ -1783.160＝1745.99

Second discriminant function F _BN =12.504X ₁ -1.212X ₂ +15.869X ₃ +115.560X ₄ +2.561X ₅ -0.026X ₆ +6.887X ₇ -1.615X ₈ +2.619X ₉ -4.518X ₁₀ +2.486X ₁₁ +5.478X ₁₂ -1860.413=1742.763

After the analysis, discrimination and result output module compares the F _BC value and the F _BN value, F _BC > F _BN , and outputs the prediction result of "the probability of the subject suffering from female breast cancer ≥ 90.0%".

The subject was diagnosed with invasive ductal carcinoma of the breast, and the clinical diagnosis results were consistent with the predicted results of the kit of the present invention.

Example 6 Using the system of Example 1 and the kit of Example 2 to predict the risk of female breast cancer in the subject

Subject: Female, 33 years old, was admitted to the Department of Breast Surgery of the Second Hospital of Jilin University, underwent breast fibroadenoma resection, fully informed the purpose and purpose of the examination, signed the informed consent form on the premise of her own voluntariness, and passed the peripheral examination. 1 mL of anticoagulant was collected intravenously.

Referring to the prediction method of Example 3, the same processing and testing were performed on the blood samples, and the results showed: L ₁ =265.37, L ₂ =276.26, L ₃ =404.10, L ₄ =404.10, L ₅ =228.72, L ₆ =230.87, L ₇ =388.82, L ₈ =388.82, L ₉ =324.57, L ₁₀ =324.57, L ₁₁ =262.90, L ₁₂ =267.41.

Calculated according to the fragment length and the following formula, denoted as X ₁ -X ₁₂ , where round represents rounding to an integer:

X ₁ =round[(L ₁ -191)/3]=25; X ₂ =round[(L ₂ -191)/3]=28;

X ₃ =round(L ₃ -379)=25; X ₄ =round(L ₄ -379)=25;

X ₅ =round[(L ₅ -202)/2]=13; X ₆ =round[(L ₆ -202)/2]=14;

X ₇ =round[(L ₇ -359)/2]=15; X ₈ =round[(L ₈ -359)/2]=15;

X ₉ =round[(L ₉ -278)/2]=23; X ₁₀ =round[(L ₁₀ -278)/2]=23;

X ₁₁ =round[(L ₁₁ -200)/4]=16; X ₁₂ =round[(L ₁₂ -200)/4]=17.

Using the computer running the system for predicting the susceptibility of female breast cancer described in Example 1, the susceptibility prediction of the subject to suffering from female breast cancer:

The age, gender, and repetition times of the short tandem sequence of the STR site are input into the system through the data input module, and the result of the discriminant function is calculated through the data calculation module:

First discriminant function F _BC =12.108X ₁ -0.933X ₂ +15.490X ₃ +112.846X ₄ +2.606X ₅ +0.178X ₆ +6.318X ₇ -1.147X ₈ +2.405X ₉ -4.504X ₁₀ +2.549X ₁₁ +5.507X ₁₂ -1783.160＝1901.877

Second discriminant function F _BN =12.504X ₁ -1.212X ₂ +15.869X ₃ +115.560X ₄ +2.561X ₅ -0.026X ₆ +6.887X ₇ -1.615X ₈ +2.619X ₉ -4.518X ₁₀ +2.486X ₁₁ +5.478X ₁₂ -1860.413＝1905.21

After the analysis, discrimination and result output module compares the F _BC value and the F _BN value, F _BC ≤ F _BN , and outputs the prediction result of "the probability of the subject not suffering from female breast cancer ≥ 77.8%".

The subject was diagnosed with breast fibroadenoma after visiting a doctor, which is a benign tumor, and the clinical diagnosis result is consistent with the prediction result of the kit of the present invention.

The above are the preferred embodiments of the present invention, and are not intended to limit the present invention. It should be pointed out that any changes, modifications, substitutions and alterations (such as: increase, decrease, change, etc.) made without departing from the principle and purpose of the present invention STR sites, using cells or tissues from other sources of the subject, using other similar statistical methods, etc.) should all be included within the scope of protection of the present invention.

SEQUENCE LISTING

<110> Jilin University

<120> A female breast cancer susceptibility prediction kit and system

<130> DI18-8180-XC47

<160> 12

<170> PatentIn version 3.3

<210> 1

<211> 18

<212> DNA

<213> Artificial

<220>

<221> primer_bind

<222> (1)..(18)

<223> STR-1 primer upstream primer

<400> 1

agggctggga agggtcta 18

<210> 2

<211> 19

<212> DNA

<213> Artificial

<220>

<221> primer_bind

<222> (1)..(19)

<223> STR-1 primer downstream primer

<400> 2

ggagaaccat cctcaccct 19

<210> 3

<211> 20

<212> DNA

<213> Artificial

<220>

<221> primer_bind

<222> (1)..(20)

<223> STR-2 primer upstream primer

<400> 3

cgcctccaag aatgtaagtg 20

<210> 4

<211> 22

<212> DNA

<213> Artificial

<220>

<221> primer_bind

<222> (1)..(22)

<223> STR-2 primer downstream primer

<400> 4

aactcaagtc tatgcttcac cc 22

<210> 5

<211> 21

<212> DNA

<213> Artificial

<220>

<221> primer_bind

<222> (1)..(21)

<223> STR-3 primer upstream primer

<400> 5

ggtttccatt gtagcatctt g 21

<210> 6

<211> 20

<212> DNA

<213> Artificial

<220>

<221> primer_bind

<222> (1)..(20)

<223> STR-3 primer downstream primer

<400> 6

gcctggttgt ttccgtagta 20

<210> 7

<211> 20

<212> DNA

<213> Artificial

<220>

<221> primer_bind

<222> (1)..(20)

<223> STR-4 primer upstream primer

<400> 7

tctgttgggt gtttgggata 20

<210> 8

<211> 20

<212> DNA

<213> Artificial

<220>

<221> primer_bind

<222> (1)..(20)

<223> STR-4 primer downstream primer

<400> 8

ttacattgtc ggtctggtcc 20

<210> 9

<211> 20

<212> DNA

<213> Artificial

<220>

<221> primer_bind

<222> (1)..(20)

<223> STR-5 primer upstream primer

<400> 9

atctcagtct ccccaagtgc 20

<210> 10

<211> 20

<212> DNA

<213> Artificial

<220>

<221> primer_bind

<222> (1)..(20)

<223> STR-5 primer downstream primer

<400> 10

tccttcaaga taaccaccga 20

<210> 11

<211> 22

<212> DNA

<213> Artificial

<220>

<221> primer_bind

<222> (1)..(22)

<223> STR-6 primer upstream primer

<400> 11

tcggttgtag gtattatcac gg 22

<210> 12

<211> 20

<212> DNA

<213> Artificial

<220>

<221> primer_bind

<222> (1)..(20)

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tgcccccatag gttttgaact 20

Claims (7)

1. A kit for predicting female breast cancer susceptibility, comprising the following components: STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer, STR-6 primer, The STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer, and STR-6 primer are respectively used to amplify the target fragment comprising the following short tandem sequences, so as to determine the following short tandem sequences Number of repetitions of the sequence: Site code starting point gene short tandem sequence STR-1 X chromosome, position 66657655 AR CAG STR-2 Chromosome 4, position 55633758 Bat-25 T STR-3 Chromosome 5, position 111646983 D5S346 GT STR-4 Chromosome 6, position 151806531 ER1 TA STR-5 Chromosome 14, position 64253561 ER2 TG STR-6 Chromosome 4, position 154587748 FGA AAAG The sequences of the STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer and STR-6 primer are as follows:

Wherein, the kit further includes: PCR amplification reaction solution, LIZ-500 molecular weight internal standard, deionized formamide and instructions for use, The instructions for use describe a method of using the kit for predicting the susceptibility of female breast cancer, which includes the following steps: (1) Extract sample DNA; (2) PCR reaction (2-1) Take out the STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer, STR-6 primer, and PCR amplification reaction solution from the refrigerator, and equilibrate to room temperature. After the components are fully dissolved, centrifuge quickly for 10 seconds; (2-2) Take 30-300ng of sample DNA, add 60μL of PCR amplification reaction solution, add deionized water to make up to 115.2μL, mix well, centrifuge quickly for 10 seconds, and then dispense the mixture at 19.2μL/well to 6 PCR reaction tubes; (2-3) Add STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer, and STR-6 primer to 0.8 μL/well of step (2-2) respectively. 6 PCR reaction tubes; cover the PCR reaction tube caps, record the sample loading situation, centrifuge quickly for 10 seconds, then transfer the PCR reaction tubes to the corresponding position of the sample tank of the PCR amplifier, record the placement sequence, and start PCR amplification Amplification reaction conditions: 95°C for 3 minutes; 95°C for 30 seconds, 60°C for 30 seconds, 72°C for 30 seconds, 10 cycles; 95°C for 30 seconds, 55°C for 30 seconds, 72°C for 30 seconds, 20 cycles ; 6 minutes at 72°C to obtain 6 sets of PCR amplification products; (3) STR fragment analysis (3-1) Take 990 μL of deionized formamide, add 10 μL of LIZ-500 molecular weight internal standard, mix well, centrifuge quickly for 10 seconds, add 10 μL/well to sequencing reaction tubes, and centrifuge quickly for 10 seconds; (3-2) Add 1 μL/well of the above-mentioned 6 sets of PCR amplification products to 6 sequencing reaction tubes, and centrifuge quickly for 10 seconds; then transfer the sequencing reaction tubes to the corresponding position of the sample tank of the PCR amplifier, 98°C Heating for 5 minutes. Immediately after the end of the program, the sequencing reaction tube was placed on the ice-water mixture and rapidly cooled to 0 °C and centrifuged rapidly for 10 seconds; then the sequencing reaction tube was transferred to the corresponding position of the sample tank of the STR locus fragment analyzer, and recorded and placed sequence, perform fragment analysis and detection; (4) Analysis and judgment of results (4-1) According to the fragment analysis results, record the fragment lengths of the two alleles of STR-1, STR-2, STR-3, STR-4, STR-5, and STR-6 respectively: The smaller fragment length value of the two alleles of STR-1 is recorded as L ₁ , and the larger fragment length value of the two alleles of STR-1 is recorded as L ₂ ; The smaller fragment length value of the two alleles of STR-2 is recorded as L ₃ , and the larger fragment length value of the two alleles of STR-2 is recorded as L ₄ ; The smaller fragment length value of the two alleles of STR-3 is recorded as L ₅ , and the larger fragment length value of the two alleles of STR-3 is recorded as L ₆ ; The smaller fragment length value of the two alleles of STR-4 is recorded as L ₇ , and the larger fragment length value of the two alleles of STR-4 is recorded as L ₈ ; The smaller fragment length value of the two alleles of STR-5 is recorded as L ₉ , and the larger fragment length value of the two alleles of STR-5 is recorded as L ₁₀ ; The smaller fragment length value of the two alleles of STR-6 is recorded as L ₁₁ , and the larger fragment length value of the two alleles of STR-6 is recorded as L ₁₂ ; (4-2) The number of repetitions of the short tandem sequence is calculated according to the fragment length and the following formula, and denoted as X ₁ -X ₁₂ , where round represents rounding to an integer: X ₁ =round[(L ₁ -191)/3]; X ₂ =round[(L ₂ -191)/3]; X ₃ =round(L ₃ -379); X ₄ =round(L ₄ -379); X ₅ =round[(L ₅ -202)/2]; X ₆ =round[(L ₆ -202)/2]; X ₇ =round[(L ₇ -359)/2]; X ₈ =round[(L ₈ -359)/2]; X ₉ =round[(L ₉ -278)/2]; X ₁₀ =round[(L ₁₀ -278)/2]; X ₁₁ =round[(L ₁₁ -200)/4]; X ₁₂ =round[(L ₁₂ -200)/4]; (4-3) Substitute the repetition times of the above short series sequence into the preset discriminant function: F _BC = 12.108X ₁ -0.933X ₂ +15.490X ₃ +112.846X ₄ +2.606X ₅ +0.178X ₆ +6.318X ₇ -1.147X ₈ +2.405X ₉ -4.504X ₁₀ +2.549X ₁₁ +5.507X ₁₂ -1783.160 F _BN = 12.504X ₁ -1.212X ₂ +15.869X ₃ +115.560X ₄ +2.561X ₅ -0.026X ₆ +6.887X ₇ -1.615X ₈ +2.619X ₉ -4.518X ₁₀ +2.486X ₁₁ +5.478X ₁₂ -1860.413; (4-4) Prediction of susceptibility to female breast cancer: Comparing the F _BC value and the F _BN value, if F _BC > F _BN , it is predicted that the probability of the subject suffering from female breast cancer is ≥90.0%; if F _BC ≤ F _BN , it is predicted that the subject will not suffer from female breast malignant tumor The odds of ≥77.8%. 2. The kit for predicting the susceptibility of female breast cancer according to claim 1, wherein the STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer, STR-6 primers were used at a concentration of 10 μM. 3. The female breast cancer susceptibility prediction kit as claimed in claim 1, wherein the PCR amplification reaction solution is the mixed solution of the following reagents: TaqDNA polymerase 5U/μL, Tris-HCl 100mM, KCl 500 mM, 0.8 vol% of ethylphenyl polyethylene glycol, 25 mM of MgCl ₂ , 10 mM of dNTP, deionized water; wherein, the pH of Tris-HCl is 8.8 at 25°C. 4 . The kit for predicting the susceptibility of female breast cancer according to claim 1 , wherein the sample is whole blood of a subject. 5 . 5. The application of the female breast cancer susceptibility prediction kit according to any one of claims 1 to 4 in the preparation of a diagnosis product for female breast cancer. 6. A system for predicting the susceptibility of female breast cancer, comprising: A device for obtaining the number of repetitions of short tandem sequences of the following STR loci in sample DNA: Site code starting point gene short tandem sequence STR-1 X chromosome, position 66657655 AR CAG STR-2 Chromosome 4, position 55633758 Bat-25 T STR-3 Chromosome 5, position 111646983 D5S346 GT STR-4 Chromosome 6, position 151806531 ER1 TA STR-5 Chromosome 14, position 64253561 ER2 TG STR-6 Chromosome 4, position 154587748 FGA AAAG ; A data processing and determination device, which includes the following modules: The data input module is used to input the age, gender, and repetition times of the short tandem sequence of the STR locus; The database management module is used for the operation management of data storage, modification, deletion, query and printing; The data calculation module is used to calculate the discriminant function result according to the repetition times of the short tandem sequence of the STR site in the data input module; The analysis and discrimination and result output module is used to compare the discriminant function results, so as to predict the susceptibility of female breast cancer, and output the results, Wherein, the number of repetitions X ₁ -X ₁₂ of the short tandem sequence of the STR site of the sample DNA is obtained by using the kit according to any one of claims 1 to 5; and in: The discriminant function includes: First discriminant function F _BC =12.108X ₁ -0.933X ₂ +15.490X ₃ +112.846X ₄ +2.606X ₅ +0.178X ₆ +6.318X ₇ -1.147X ₈ +2.405X ₉ -4.504X ₁₀ +2.549X ₁₁ +5.507X ₁₂ -1783.160 Second discriminant function F _BN =12.504X ₁ -1.212X ₂ +15.869X ₃ +115.560X ₄ +2.561X ₅ -0.026X ₆ +6.887X ₇ -1.615X ₈ +2.619X ₉ -4.518X ₁₀ +2.486X ₁₁ +5.478X ₁₂ -1860.413 In the discriminant function, X ₁ is the number of repetitions of the short tandem sequence of the smaller fragment in the two alleles of STR-1; X ₂ is the number of repetitions of the short tandem sequence of the larger fragment in the two alleles of STR-1; X ₃ is the number of repetitions of the short tandem sequence of the smaller fragment in the two alleles of STR-2; X ₄ is the number of repetitions of the short tandem sequence of the larger fragment in the two alleles of STR-2; X ₅ is the number of repetitions of the short tandem sequence of the smaller fragment in the two alleles of STR-3; X ₆ is the number of repetitions of the short tandem sequence of the larger fragment in the two alleles of STR-3; X ₇ is the number of repetitions of the short tandem sequence of the smaller fragment in the two alleles of STR-4; X ₈ is the number of repetitions of the short tandem sequence of the larger fragment in the two alleles of STR-4; X ₉ is the number of repetitions of the short tandem sequence of the smaller fragment in the two alleles of STR-5; X ₁₀ is the number of repetitions of the short tandem sequence of the larger fragment in the two alleles of STR-5; X ₁₁ is the number of repetitions of the short tandem sequence of the smaller fragment in the two alleles of STR-6; X ₁₂ is the number of repetitions of the short tandem sequence of the larger fragment in the two alleles of STR-6; Among them, X ₁ ^- X ₁₂ are calculated according to the length of the segment and as follows, where round represents rounding to an integer: X ₁ =round[(L ₁ -191)/3]; X ₂ =round[(L ₂ -191)/3]; X ₃ =round(L ₃ -379); X ₄ =round(L ₄ -379); X ₅ =round[(L ₅ -202)/2]; X ₆ =round[(L ₆ -202)/2]; X ₇ =round[(L ₇ -359)/2]; X ₈ =round[(L ₈ -359)/2]; X ₉ =round[(L ₉ -278)/2]; X ₁₀ =round[(L ₁₀ -278)/2]; X ₁₁ =round[(L ₁₁ -200)/4]; X ₁₂ =round[(L ₁₂ -200)/4]; In X ₁ ^- X ₁₂ , L ₁ is the smaller fragment length value of the two alleles of STR-1, and L ₂ is the larger fragment length value of the two alleles of STR-1; L ₃ is the smaller fragment length value of the two alleles of STR-2, and L ₄ is the larger fragment length value of the two alleles of STR-2; L ₅ is the smaller fragment length value of the two alleles of STR-3, and L ₆ is the larger fragment length value of the two alleles of STR-3; L ₇ is the smaller fragment length value of the two alleles of STR-4, and L ₈ is the larger fragment length value of the two alleles of STR-4; L ₉ is the smaller fragment length value of the two alleles of STR-5, and L ₁₀ is the larger fragment length value of the two alleles of STR-5; L ₁₁ is the smaller fragment length value of the two alleles of STR-6, and L ₁₂ is the larger fragment length value of the two alleles of STR-6; The analysis, discrimination and result output module compares the calculation result of the first discriminant function F _BC with the calculation result of the second discriminant function F _BN , and if F _BC > _{FB BN} , it outputs “the subject suffers from a female breast cancer. If F _BC ≤ F _BN , output the prediction result of "the probability of the subject not suffering from female breast cancer ≥ 77.8%". 7. The application of the system for predicting the susceptibility of female breast malignant tumor according to claim 6 in the preparation of a female breast malignant tumor prediction product or a female breast malignant tumor diagnosis product.

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