CN108866190A - A kind of malignant tumor of ovary neurological susceptibility prediction kit and system - Google Patents

A kind of malignant tumor of ovary neurological susceptibility prediction kit and system Download PDF

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CN108866190A
CN108866190A CN201810766322.5A CN201810766322A CN108866190A CN 108866190 A CN108866190 A CN 108866190A CN 201810766322 A CN201810766322 A CN 201810766322A CN 108866190 A CN108866190 A CN 108866190A
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任明
郝书弘
王晓峰
杨麒巍
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Jilin University
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Abstract

The present invention relates to a kind of malignant tumor of ovary neurological susceptibilities to predict kit comprising following components:STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer, STR-6 primer;Further, may also include:Pcr amplification reaction liquid, LIZ-500 molecular weight internal standard, deionized formamide.Malignant tumor of ovary neurological susceptibility prediction kit of the present invention can be used for the diagnosis and neurological susceptibility prediction of malignant tumor of ovary.The present invention also provides a kind of malignant tumor of ovary neurological susceptibility forecasting systems.

Description

Ovarian malignant tumor susceptibility prediction kit and system
Technical Field
The present invention relates to the field of biomedicine. In particular to a kit for predicting susceptibility of ovarian malignant tumor and a system for predicting susceptibility of ovarian malignant tumor. More specifically, the invention relates to a kit for detecting STR of ovarian malignant tumor susceptibility related genes by Short Tandem Repeat (STR) locus fragment analysis method, and early warning is carried out on ovarian malignant tumor susceptibility of a detected object by combining with a discriminant analysis statistical method.
Background
The tumor is a disease closely related to genetic genes, the molecular genetics basis of the tumor is researched, and further a tumor specific genetics marker is provided, so that the tumor specific genetics marker is expected to provide a simple and feasible method for common detection, clinical diagnosis, personalized treatment, disease tracking after recovery and the like. However, the individual differences of patients, the intercrossing of related biomolecular events at different stages of development, etc. all bring great difficulties to the work.
Ovarian malignancies are one of the most common malignancies of female reproductive organs, with the third highest incidence next to cervical and uterine body cancers. However, the death rate of ovarian epithelial cancer accounts for the first position of various gynecological tumors, and the life of women is seriously threatened. Because the embryonic development, tissue dissection and endocrine function of the ovary are complex and the early symptoms are not typical, the identification of the tissue type and the benign and malignant properties of the ovarian tumor before operation is quite difficult. Among ovarian malignancies, the epithelial cancers are most common, followed by malignant germ cell tumors. The tumor is only limited to 30 percent of the ovary in the operation of patients with the ovarian epithelial cancer, and most of the tumor has spread to the organs of the uterus, bilateral appendages, the omentum majus and the pelvic cavity, so the early diagnosis is a big problem. Therefore, the method can predict the susceptibility of ovarian malignant tumor of the human population to be detected, and is beneficial to improving the risk awareness of the disease and finding and treating the disease as soon as possible.
A large number of studies have shown that genetic polymorphisms of tumor-associated genes play a key role in the development of malignant tumors. However, the development of tumors is a very complex process, and the diagnosis of the disease using changes in a single molecular genetic marker is clearly impossible and not scientific. In the prior art, accurate early warning on tumor susceptibility cannot be performed only through genetic information, and the current early identification and prediction method for tumors needs to be improved. The invention relates to a kit for performing early warning on susceptibility of ovarian malignant tumor by jointly detecting a plurality of STR loci with high relevance to occurrence of ovarian malignant tumor through an STR locus fragment analysis method and combining a discriminant analysis statistical method.
Disclosure of Invention
In order to solve the problems in the prior art, the invention relates to a method for jointly detecting a plurality of STR loci with high relevance to the occurrence of ovarian malignant tumor by an STR locus fragment analysis method, and early warning is carried out on the susceptibility of the ovarian malignant tumor by combining a discriminant analysis statistical method.
The present invention has been completed based on the following findings of the inventors: the inventor finds that the repetition times of short tandem sequences of each independent STR locus has no significant correlation with the ovarian malignant tumor of the detected object, and the combination of the repetition times of the short tandem sequences of certain specific STR loci has close relation with the ovarian malignant tumor of the detected object by analyzing STRs of the genomic DNA of the ovarian malignant tumor detected object and a healthy control detected object and verifying a large number of ovarian malignant tumor samples and control samples.
To this end, the present invention proposes a set of isolated STR loci that have a high association with the development of ovarian malignancies. According to an embodiment of the present invention, these isolated STR loci comprise the nucleotide sequences shown as STR-1 to STR-6 (Table 1). By using these isolated STR loci as references, susceptibility to ovarian malignancy can be predicted efficiently.
TABLE 1
Locus code Starting position Belonging 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 above detailed description of STR sites, those skilled in the art can log in relevant databases (such as GeneBank, Nucleotide, etc.) to obtain the details, which are not described herein. The inventor surprisingly finds that the ovarian malignant tumor susceptibility can be early warned by a statistical analysis method, such as analyzing the cell genome of a detected object to obtain the repetition times of the short tandem sequence of each STR locus and carrying out discriminant analysis by taking the repetition times as independent variables.
On the basis, one of the technical problems solved by the invention is to provide a kit for predicting susceptibility to ovarian malignant tumor, which comprises the following components: STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer and STR-6 primer, wherein the primers are respectively used for amplifying target fragments containing short tandem sequences listed in Table 1 so as to determine the repetition times of the short tandem sequences.
Preferably, the ovarian malignancy susceptibility prediction kit of the present invention further comprises: PCR amplification reaction liquid, LIZ-500 molecular weight internal standard and deionized formamide.
In the kit for predicting susceptibility to ovarian malignant tumor of the present invention, preferably, 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 shown in table 2 below, and more preferably, the concentrations of the primers are all 10 μ M:
TABLE 2
In table 2, HEX, FAM, and ROX are all fluorophores labeling the 5' end, HEX is hexachloro-6-methylfluorescein, FAM is 6-carboxyfluorescein, and ROX is ROX reference dye.
In the kit for predicting susceptibility to ovarian malignant tumor of the present invention, preferably, the PCR amplification reaction solution is a mixture of the following reagents: TaqDNA polymerase (5U/. mu.L), Tris-HCl (100mM, pH 8.8 at 25 ℃), KCl (500mM), ethylphenylpolyethyleneglycol (0.8% (v/v)), MgCl2(25mM), dNTP (10mM), deionized water.
More preferably, the PCR amplification reaction solution is stored at-20 ℃.
In the kit for predicting susceptibility to ovarian malignant tumor of the invention, preferably, the LIZ-500 molecular weight internal standard can be stored at-20 ℃;
in the kit for predicting susceptibility to ovarian malignant tumor of the present invention, preferably, the deionized formamide can be stored at 2-8 ℃.
Preferably, the kit for predicting susceptibility to ovarian malignancy further comprises instructions for use.
The application instruction describes a use method of the ovarian malignant tumor susceptibility prediction kit, which comprises the following steps:
(1) extracting sample DNA;
(2) PCR reaction
(2-1) taking out the STR-1 primer, the STR-2 primer, the STR-3 primer, the STR-4 primer, the STR-5 primer, the STR-6 primer and the PCR amplification reaction solution from a refrigerator, balancing to room temperature, fully dissolving each component, and respectively and rapidly centrifuging for 10 seconds;
(2-2) adding 30-300ng of sample DNA into 60 mu L of PCR amplification reaction solution, adding deionized water to supplement to 115.2 mu L, fully and uniformly mixing, quickly centrifuging for 10 seconds, and subpackaging the mixed solution into 6 PCR reaction tubes according to 19.6 mu L/hole;
(2-3) respectively adding an STR-1 primer, an STR-2 primer, an STR-3 primer, an STR-4 primer, an STR-5 primer and an STR-6 primer into the 6 PCR reaction tubes in the step (2-2) according to 0.8 mu L/hole; covering a PCR reaction tube cover, recording the sample adding condition, quickly centrifuging for 10 seconds, then transferring the PCR reaction tube to a corresponding position of a sample groove of a PCR amplification instrument, recording the placing sequence, and starting the PCR amplification reaction; the amplification reaction conditions are as follows: 3 minutes at 95 ℃; 30 seconds at 95 ℃, 30 seconds at 60 ℃ and 30 seconds at 72 ℃ for 10 cycles; 30 seconds at 95 ℃, 30 seconds at 55 ℃, 30 seconds at 72 ℃ and 20 cycles; 6 groups of PCR amplification products are obtained at 72 ℃ for 6 minutes;
(3) STR fragment analysis
(3-1) adding 990 mu L of deionized formamide into 10 mu L of LIZ-500 molecular weight internal standard, fully and uniformly mixing, quickly centrifuging for 10 seconds, respectively adding into a sequencing reaction tube according to 10 mu L/hole, and quickly centrifuging for 10 seconds;
(3-2) adding the 6 groups of PCR amplification products into 6 sequencing reaction tubes according to 1 mu L/hole respectively, and quickly centrifuging for 10 seconds; then transferring the sequencing reaction tube to a corresponding position of a sample tank of a PCR (polymerase chain reaction) amplification instrument, heating at 98 ℃ for 5 minutes, immediately placing the sequencing reaction tube on an ice-water mixture after the program is finished, rapidly cooling to 0 ℃, and rapidly centrifuging for 10 seconds; then transferring the sequencing reaction tube to a corresponding position of a sample groove of an STR locus fragment analyzer, recording the placement sequence, and performing fragment analysis detection;
(4) analysis and determination of results
(4-1) respectively recording the fragment lengths of two alleles at each site of STR-1, STR-2, STR-3, STR-4, STR-5 and STR-6 according to the fragment analysis result:
the length of the smaller of the two STR-1 alleles is recorded as L1And the length of the larger fragment of the two STR-1 alleles is designated as L2
The length of the smaller fragment of the two STR-2 alleles is recorded as L3And the length of the larger fragment of the two STR-2 alleles is designated as L4
The length of the smaller fragment of the two STR-3 alleles is recorded as L5And the length of the larger fragment of the two STR-3 alleles is marked as L6
The length of the smaller fragment of the two STR-4 alleles is recorded as L7And the length of the larger fragment of the two STR-4 alleles is marked as L8
The length of the smaller of the two STR-5 alleles is recorded as L9And the length of the larger fragment of the two STR-5 alleles is designated as L10
The length of the smaller fragment of the two STR-6 alleles was designated L11And the length of the larger fragment of the two STR-6 alleles is marked as L12
(4-2) the number of repetitions of the short tandem sequence is calculated from the fragment length and the following formula, and is denoted as X1-X12Where round stands for rounded integer:
X1=round[(L1-191)/3];X2=round[(L2-191)/3];
X3=round(L3-379);X4=round(L4-379);
X5=round[(L5-202)/2];X6=round[(L6-202)/2];
X7=round[(L7-359)/2];X8=round[(L8-359)/2];
X9=round[(L9-278)/2];X10=round[(L10-278)/2];
X11=round[(L11-200)/4];X12=round[(L12-200)/4];
(4-3) substituting the number of the short tandem sequence repetitions into a preset discriminant function:
FOC=7.213 X1+5.611 X2+8.722 X3+102.277 X4+3.013 X5-1.076 X6-3.097 X7+3.897X8+0.695 X9-0.691 X10+0.914 X11-3.943 X12-1530.478
FON=7.556 X1+4.831 X2+8.999 X3+102.984 X4+2.680 X5-0.960 X6-2.482 X7+3.491X8+0.857 X9-0.690 X10+1.735 X11-4.203 X12-1550.198
(4-4) prediction of susceptibility to ovarian malignancy:
comparison FOCValue sum FONValue if FOC>FONPredicting the probability of the tested object suffering from ovarian malignant tumor to be more than or equal to 88.9 percent; if FOC≤FONAnd predicting that the probability that the detected object does not suffer from the ovarian malignant tumor is more than or equal to 83.3 percent.
In the present invention,
preferably, the sample DNA extracted in step (1) can be performed using a commercially available genomic DNA extraction kit according to the kit instructions. The sample may be whole blood of a subject.
Preferably, the rotational speed of all the centrifuges in the method of use is preferably 3000 g/min.
The probability of suffering from ovarian malignant tumor in the invention is the sum of the probability of suffering from ovarian malignant tumor already and the probability of suffering from ovarian malignant tumor in the future. Therefore, the kit can be used for diagnosing ovarian malignant tumor; the method can also be used for risk early warning of future ovarian malignant tumors, can assist a detected object in risk prevention, and reduces the incidence of the ovarian malignant tumors by means of medicine conditioning, change of daily work and rest, diet rules, regular physical examination and the like.
The second technical problem solved by the invention is to provide a method for predicting susceptibility to ovarian malignant tumor, namely, the kit is used and the method is operated according to the instruction.
The invention solves the third technical problem by providing the application of the ovarian malignant tumor susceptibility prediction kit in preparing ovarian malignant tumor diagnosis products.
The fourth technical problem to be solved by the present invention is to provide a system for predicting susceptibility to ovarian malignant tumor, which comprises:
a device for obtaining the repeat times of the STR locus short tandem sequence of the sample DNA;
data processing and decision device, comprising the following modules:
the data input module is used for inputting the age, the sex and the STR locus short tandem sequence repetition times of the detected object;
the database management module is used for the operation management of data storage, modification, deletion, inquiry and printing;
the data calculation module is used for calculating a discrimination function result according to the repeat times of the STR locus short serial sequence in the data input module;
and the analysis, discrimination and result output module is used for comparing the discrimination function results so as to predict the susceptibility of the ovarian malignant tumor and output the result.
Wherein,
the number of times of the STR locus short tandem sequence repetition is 6 pairs of the number of times of the STR locus short tandem sequence repetition:
locus code Starting position Belonging 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 FOC=7.213 X1+5.611 X2+8.722 X3+102.277 X4+3.013 X5-1.076X6-3.097 X7+3.897 X8+0.695 X9-0.691 X10+0.914 X11-3.943 X12-1530.478
Second discrimination function FON=7.556 X1+4.831 X2+8.999 X3+102.984 X4+2.680 X5-0.960X6-2.482 X7+3.491 X8+0.857 X9-0.690 X10+1.735 X11-4.203 X12-1550.198
In the case of the discriminant function,
X1the number of repeats of the short tandem sequence for the smaller of the two alleles of STR-1;
X2the number of repeats of the short tandem sequence for the larger of the two alleles of STR-1;
X3the number of repeats of the short tandem sequence for the smaller of the two alleles of STR-2;
X4the number of repeats of the short tandem sequence for the larger of the two alleles of STR-2;
X5the number of repeats of the short tandem sequence for the smaller of the two alleles of STR-3;
X6the number of repeats of the short tandem sequence for the larger of the two alleles of STR-3;
X7the number of repeats of the short tandem sequence for the smaller of the two alleles of STR-4;
X8the number of repeats of the short tandem sequence for the larger of the two alleles of STR-4;
X9the number of repeats of the short tandem sequence for the smaller of the two alleles of STR-5;
X10the number of repeats of the short tandem sequence for the larger of the two alleles of STR-5;
X11the number of repeats of the short tandem sequence for the smaller of the two alleles of STR-6;
X12the number of repeats of the short tandem sequence for the larger of the two alleles of STR-6;
wherein, X1-X12Calculated from the fragment length and the following formula, where round stands for rounded integer:
X1=round[(L1-191)/3];X2=round[(L2-191)/3];
X3=round(L3-379);X4=round(L4-379);
X5=round[(L5-202)/2];X6=round[(L6-202)/2];
X7=round[(L7-359)/2];X8=round[(L8-359)/2];
X9=round[(L9-278)/2];X10=round[(L10-278)/2];
X11=round[(L11-200)/4];X12=round[(L12-200)/4];
X1-X12in, L1Is the smaller segment length value, L, of the two alleles of STR-12Is the larger fragment length value of the two alleles of STR-1;
L3is the smaller segment length value, L, of the two alleles of STR-24Is STR-2 two allelic groupsDue to the larger segment length value;
L5is the smaller segment length value, L, of the two alleles of STR-36Is the larger fragment length value of the two alleles of STR-3;
L7is the smaller segment length value, L, of the two alleles of STR-48Is the larger fragment length value of the two alleles of STR-4;
L9is the smaller segment length value, L, of the two alleles of STR-510Is the larger fragment length value of the two alleles of STR-5;
L11is the smaller segment length value, L, of the two STR-6 alleles12Is the larger fragment length value of the two alleles of STR-6;
the analysis discrimination and result output module outputs a first discrimination function FOCAnd a second discrimination function FONIf F is the result of the calculation ofOC>FONOutputting a prediction result that the probability of the ovarian malignant tumor of the detected object is more than or equal to 88.9%; if FOC≤FONAnd outputting a prediction result that the probability that the detected object does not suffer from ovarian malignant tumor is more than or equal to 83.3 percent.
The device for obtaining the repetition times of the STR locus short tandem sequence of the sample DNA can comprise an STR locus fragment analyzer, a PCR amplification instrument and the like; the data processing and determining device may be a computer or the like.
The fifth technical problem to be solved by the invention is to provide the application of the ovarian malignant tumor susceptibility prediction system in the preparation of ovarian malignant tumor prediction products, ovarian malignant tumor diagnosis products and ovarian health auxiliary products.
The sixth technical problem to be solved by the invention is to provide an ovarian malignant tumor prediction product, an ovarian malignant tumor diagnosis product or an ovarian health auxiliary product, which comprises the ovarian malignant tumor susceptibility prediction system.
The test material used in the present invention is human genomic DNA, which theoretically does not change during the life of a human. The human genome DNA encodes all life activities of human, so theoretically, the risk of a detected object suffering from a certain disease can be predicted at an early stage by detecting the genome DNA, and even the detected object can be predicted at birth.
The development of tumors is a very complex process. The molecular genetics basis of tumor research is expected to provide a simple and feasible method for common detection, clinical diagnosis, personalized treatment, disease tracking after recovery and the like. However, the individual differences of patients, the intercrossing of related biomolecular events at different stages of development, etc. all bring great difficulties to the work. It is clearly impossible and not scientific to use single molecular genetic changes to diagnose tumors. The inventor applies modern molecular biology technology to carry out combined analysis on a plurality of STRs of genomic DNA of a detected object, and combines statistical analysis methods such as discriminant analysis and the like, thereby inventing a kit for early warning of susceptibility to ovarian malignant tumor.
Drawings
FIG. 1 is a schematic diagram of modules included in a data processing and determining device of the ovarian malignancy susceptibility predicting system according to the present invention.
Detailed Description
The invention will be better understood from the following description of specific embodiments thereof, taken in conjunction with the accompanying drawings and examples. The experimental procedures used in the following examples are all conventional procedures unless otherwise specified. Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the accompanying drawings are illustrative only for the purpose of explaining the present invention, and are not to be construed as limiting the present invention.
The PCR amplification apparatus in the examples was a Mastercycler nexus amplification apparatus (purchased from eppendorf, USA);
the STR locus fragment analyzer in the examples was a 3730XL sequencing analyzer (purchased from ABI, usa);
the DNA extraction kit in the examples was a blood DNAout kit (purchased from engze, beijing);
the rotational speed of all the centrifuges in the examples was 3000 g/min.
Example 1 ovarian malignancy susceptibility prediction system
A ovarian malignancy susceptibility prediction system, comprising:
a device for obtaining the repeat times of the STR locus short tandem sequence of the sample DNA;
data processing and decision device, comprising the following modules (fig. 1):
the data input module is used for inputting the age, the sex and the STR locus short tandem sequence repetition times of the detected object;
the database management module is used for the operation management of data storage, modification, deletion, inquiry and printing;
the data calculation module is used for calculating a discrimination function result according to the repeat times of the STR locus short serial sequence in the data input module;
and the analysis, discrimination and result output module is used for comparing the discrimination function results so as to predict the susceptibility of the ovarian malignant tumor and output the result.
Wherein,
the number of times of the STR locus short tandem sequence repetition is 6 pairs of the number of times of the STR locus short tandem sequence repetition:
locus code Starting position Belonging 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 FOC=7.213 X1+5.611 X2+8.722 X3+102.277 X4+3.013 X5-1.076X6-3.097 X7+3.897 X8+0.695 X9-0.691 X10+0.914 X11-3.943 X12-1530.478
Second discrimination function FON=7.556 X1+4.831 X2+8.999 X3+102.984 X4+2.680 X5-0.960X6-2.482 X7+3.491 X8+0.857 X9-0.690 X10+1.735 X11-4.203 X12-1550.198
In the case of the discriminant function,
X1the number of repeats of the short tandem sequence for the smaller of the two alleles of STR-1;
X2the number of repeats of the short tandem sequence for the larger of the two alleles of STR-1;
X3the number of repeats of the short tandem sequence for the smaller of the two alleles of STR-2;
X4the number of repeats of the short tandem sequence for the larger of the two alleles of STR-2;
X5the number of repeats of the short tandem sequence for the smaller of the two alleles of STR-3;
X6the number of repeats of the short tandem sequence for the larger of the two alleles of STR-3;
X7the number of repeats of the short tandem sequence for the smaller of the two alleles of STR-4;
X8the number of repeats of the short tandem sequence for the larger of the two alleles of STR-4;
X9the number of repeats of the short tandem sequence for the smaller of the two alleles of STR-5;
X10the number of repeats of the short tandem sequence for the larger of the two alleles of STR-5;
X11the number of repeats of the short tandem sequence for the smaller of the two alleles of STR-6;
X12the number of repeats of the short tandem sequence for the larger fragment of both alleles of STR-6.
Wherein, X1-X12Calculated from the fragment length and the following formula, where round stands for rounded integer:
X1=round[(L1-191)/3];X2=round[(L2-191)/3];
X3=round(L3-379);X4=round(L4-379);
X5=round[(L5-202)/2];X6=round[(L6-202)/2];
X7=round[(L7-359)/2];X8=round[(L8-359)/2];
X9=round[(L9-278)/2];X10=round[(L10-278)/2];
X11=round[(L11-200)/4];X12=round[(L12-200)/4];
X1-X12in, L1Is the smaller segment length value, L, of the two alleles of STR-12Is the larger fragment length value of the two alleles of STR-1;
L3is the smaller segment length value, L, of the two alleles of STR-24Is the larger fragment length value of the two alleles of STR-2;
L5is the smaller segment length value, L, of the two alleles of STR-36Is the larger fragment length value of the two alleles of STR-3;
L7is the smaller segment length value, L, of the two alleles of STR-48Is the larger fragment length value of the two alleles of STR-4;
L9is the smaller segment length value, L, of the two alleles of STR-510Is the larger fragment length value of the two alleles of STR-5;
L11is the smaller segment length value, L, of the two STR-6 alleles12Is the larger fragment length value of the two alleles of STR-6;
the analysis discrimination and result output module outputs a first discrimination function FOCAnd a second discrimination function FONThe results of the calculations of (a) are compared,if FOC>FONOutputting a prediction result that the probability of the ovarian malignant tumor of the detected object is more than or equal to 88.9%; if FOC≤FONAnd outputting a prediction result that the probability that the detected object does not suffer from ovarian malignant tumor is more than or equal to 83.3 percent.
Example 2 ovarian malignant tumor susceptibility prediction kit
A kit for predicting susceptibility to ovarian malignancy, comprising the following components: STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer, STR-6 primer, PCR amplification reaction liquid, LIZ-500 molecular weight internal standard, deionized formamide and an instruction book.
The concentrations of the STR-1 primer, the STR-2 primer, the STR-3 primer, the STR-4 primer, the STR-5 primer and the STR-6 primer are all 10 mu M, and the primer sequences are shown in the following table:
the PCR amplification reaction solution is a mixed solution of the following reagents: TaqDNA polymerase (5U/. mu.L), Tris-HCl (100mM, pH 8.8 at 25 ℃), KCl (500mM), ethylphenylpolyethyleneglycol (0.8% (v/v)), MgCl2(25mM), dNTP (10mM), deionized water.
Storing the PCR amplification reaction solution at-20 ℃; LIZ-500 molecular weight internal standard is preserved at-20 ℃; storing deionized formamide at 2-8 deg.C.
The kit further comprises instructions for use.
Example 3 Using the System of example 1 and the kit of example 2 to predict the risk of ovarian malignancy in a subject
The detected object is: female, age 58, visiting the department of obstetrics and gynecology of the second hospital of Jilin university, with sufficient informed examination purpose and use, signed an informed consent and collected 1mL of anticoagulated blood via the peripheral vein on the premise of his own accord.
The following procedure was carried out using the kit of example 2 according to the method described in the kit instructions:
(1) extracting sample DNA: extracting blood genome DNA by using a DNA extraction kit;
(2) PCR reaction
(2-1) taking out the STR-1 primer, the STR-2 primer, the STR-3 primer, the STR-4 primer, the STR-5 primer, the STR-6 primer and the PCR amplification reaction solution from a refrigerator, balancing to room temperature, fully dissolving each component, and respectively and rapidly centrifuging for 10 seconds;
(2-2) adding 100ng of sample DNA into 60 mu L of PCR amplification reaction solution, adding deionized water to supplement to 115.2 mu L, fully and uniformly mixing, quickly centrifuging for 10 seconds, and subpackaging the mixed solution into 6 PCR reaction tubes according to 19.6 mu L/hole;
(2-3) respectively adding an STR-1 primer, an STR-2 primer, an STR-3 primer, an STR-4 primer, an STR-5 primer and an STR-6 primer into the 6 PCR reaction tubes in the step (2-2) according to 0.8 mu L/hole; covering a PCR reaction tube cover, recording the sample adding condition, quickly centrifuging for 10 seconds, then transferring the PCR reaction tube to a corresponding position of a sample groove of a PCR amplification instrument, recording the placing sequence, and starting the PCR amplification reaction; the amplification reaction conditions are as follows: 3 minutes at 95 ℃; 30 seconds at 95 ℃, 30 seconds at 60 ℃ and 30 seconds at 72 ℃ for 10 cycles; 30 seconds at 95 ℃, 30 seconds at 55 ℃, 30 seconds at 72 ℃ and 20 cycles; 6 groups of PCR amplification products are obtained at 72 ℃ for 6 minutes;
(3) STR fragment analysis
(3-1) adding 990 mu L of deionized formamide into 10 mu L of LIZ-500 molecular weight internal standard, fully and uniformly mixing, quickly centrifuging for 10 seconds, respectively adding into a sequencing reaction tube according to 10 mu L/hole, and quickly centrifuging for 10 seconds;
(3-2) adding the 6 groups of PCR amplification products into 6 sequencing reaction tubes according to 1 mu L/hole respectively, and quickly centrifuging for 10 seconds; then transferring the sequencing reaction tube to a corresponding position of a sample tank of a PCR (polymerase chain reaction) amplification instrument, heating at 98 ℃ for 5 minutes, immediately placing the sequencing reaction tube on an ice-water mixture after the program is finished, rapidly cooling to 0 ℃, and rapidly centrifuging for 10 seconds; then transferring the sequencing reaction tube to a corresponding position of a sample groove of an STR locus fragment analyzer, recording the placement sequence, and performing fragment analysis detection;
(4) analysis and determination of results
(4-1) respectively recording the fragment lengths of two alleles at each site of STR-1, STR-2, STR-3, STR-4, STR-5 and STR-6 according to the fragment analysis result: the length of the smaller of the two STR-1 alleles is recorded as L1And the length of the larger fragment of the two STR-1 alleles is designated as L2(ii) a The length of the smaller fragment of the two STR-2 alleles is recorded as L3And the length of the larger fragment of the two STR-2 alleles is designated as L4(ii) a The length of the smaller fragment of the two STR-3 alleles is recorded as L5And the length of the larger fragment of the two STR-3 alleles is marked as L6(ii) a The length of the smaller fragment of the two STR-4 alleles is recorded as L7And the length of the larger fragment of the two STR-4 alleles is marked as L8(ii) a The length of the smaller of the two STR-5 alleles is recorded as L9And the length of the larger fragment of the two STR-5 alleles is designated as L10(ii) a The length of the smaller fragment of the two STR-6 alleles was designated L11And the length of the larger fragment of the two STR-6 alleles is marked as L12(ii) a The results show that: l is1=268.74,L2=279.73,L3=403.17,L4=403.17,L5=228.53,L6=228.53,L7=382.15,L8=401.6,L9=309.62,L10=318.17,L11=249.36,L12=256.62。
(4-2) according to fragment lengthDegree and the following formula, and is recorded as X1-X12Where round stands for rounded integer:
X1=round[(L1-191)/3]=26;X2=round[(L2-191)/3]=30;
X3=round(L3-379)=24;X4=round(L4-379)=24;
X5=round[(L5-202)/2]=13;X6=round[(L6-202)/2]=13;
X7=round[(L7-359)/2]=12;X8=round[(L8-359)/2]=21;
X9=round[(L9-278)/2]=16;X10=round[(L10-278)/2]=20;
X11=round[(L11-200)/4]=12;X12=round[(L12-200)/4]=14。
(4-3) using a computer running the system for predicting susceptibility to ovarian malignancy described in example 1, predicting susceptibility to ovarian malignancy in a subject:
inputting the age, the sex and the STR locus short tandem sequence repetition times of the detected object into a system through a data input module, and calculating the result of a discriminant function through a data calculation module:
first discriminant function FOC=7.213 X1+5.611 X2+8.722 X3+102.277 X4+3.013 X5-1.076X6-3.097 X7+3.897 X8+0.695 X9-0.691 X10+0.914 X11-3.943 X12-1530.478=1512.286
Second discrimination function FON=7.556 X1+4.831 X2+8.999 X3+102.984 X4+2.680 X5-0.960X6-2.482 X7+3.491 X8+0.857 X9-0.690 X10+1.735 X11-4.203 X12-1550.198=1506.557
Analyzed, determined and result output module compared FOCValue sum FONValue, FOC>FONAnd outputting a prediction result that the probability of the ovarian malignant tumor of the detected object is more than or equal to 88.9 percent.
The examinee is diagnosed as ovarian high-grade serous adenocarcinoma by a pathological examination after the examination of laparotomy, and the clinical diagnosis result of the examinee is consistent with the prediction result of the kit.
Example 4 Using the System of example 1 and the kit of example 2 to predict the risk of ovarian malignancy in a subject
The detected object is: woman, age 76, visiting the department of obstetrics and gynecology of the second hospital of Jilin university, with sufficient informed examination purposes and uses, signed an informed consent and collected 1mL of anticoagulated blood via the peripheral vein on the premise of his own accord.
The same treatments and tests were carried out on blood samples, with reference to the prediction method of example 3, and the results show that: l is1=260.10,L2=260.10,L3=404.22,L4=404.22,L5=228.59,L6=228.59,L7=385.03,L8=388.90,L9=311.78,L10=324.52,L11=259.24,L12=263.80。
Calculated according to the fragment length and the following formula, denoted X1-X12Where round stands for rounded integer:
X1=round[(L1-191)/3]=23;X2=round[(L2-191)/3]=23;
X3=round(L3-379)=25;X4=round(L4-379)=25;
X5=round[(L5-202)/2]=13;X6=round[(L6-202)/2]=13;
X7=round[(L7-359)/2]=13;X8=round[(L8-359)/2]=15;
X9=round[(L9-278)/2]=17;X10=round[(L10-278)/2]=23;
X11=round[(L11-200)/4]=15;X12=round[(L12-200)/4]=16。
using a computer running the system for predicting susceptibility to ovarian malignancy described in example 1, a susceptibility to developing ovarian malignancy in a subject is predicted by:
inputting the age, the sex and the STR locus short tandem sequence repetition times of the detected object into a system through a data input module, and calculating the result of a discriminant function through a data calculation module:
first discriminant function FOC=7.213 X1+5.611 X2+8.722 X3+102.277 X4+3.013 X5-1.076X6-3.097 X7+3.897 X8+0.695 X9-0.691 X10+0.914 X11-3.943 X12-1530.478=1529.368
Second discrimination function FON=7.556 X1+4.831 X2+8.999 X3+102.984 X4+2.680 X5-0.960X6-2.482 X7+3.491 X8+0.857 X9-0.690 X10+1.735 X11-4.203 X12-1550.198=1534.213
Analyzed, determined and result output module compared FOCValue sum FONValue, FOC≤FONOutputting that the subject does not suffer from ovarian malignancyThe prediction result that the probability of the tumor is more than or equal to 83.3 percent.
The detected object is diagnosed as urethritis after the visit, and the clinical diagnosis result of the detected object is consistent with the prediction result of the kit.
Example 5 Using the System of example 1 and the kit of example 2 to predict the risk of ovarian malignancy in a subject
The detected object is: women, age 63, visiting the department of obstetrics and gynecology of the second hospital of Jilin university, performing laparotomy, with sufficient informed examination purpose and use, under his voluntary premise, signed an informed consent, and collected 1mL of anticoagulated blood via the peripheral vein.
The same treatments and tests were carried out on blood samples, with reference to the prediction method of example 3, and the results show that: l is1=260.38,L2=273.9,L3=403.14,L4=403.14,L5=244.15,L6=246.34,L7=382.95,L8=398.47,L9=309.99,L10=312.14,L11=251.52,L12=253.36。
Calculated according to the fragment length and the following formula, denoted X1-X12Where round stands for rounded integer:
X1=round[(L1-191)/3]=23;X2=round[(L2-191)/3]=28;
X3=round(L3-379)=24;X4=round(L4-379)=24;
X5=round[(L5-202)/2]=21;X6=round[(L6-202)/2]=22;
X7=round[(L7-359)/2]=12;X8=round[(L8-359)/2]=20;
X9=round[(L9-278)/2]=16;X10=round[(L10-278)/2]=17;
X11=round[(L11-200)/4]=13;X12=round[(L12-200)/4]=13。
using a computer running the system for predicting susceptibility to ovarian malignancy described in example 1, a susceptibility to developing ovarian malignancy in a subject is predicted by:
inputting the age, the sex and the STR locus short tandem sequence repetition times of the detected object into a system through a data input module, and calculating the result of a discriminant function through a data calculation module:
first discriminant function FOC=7.213 X1+5.611 X2+8.722 X3+102.277 X4+3.013 X5-1.076X6-3.097 X7+3.897 X8+0.695 X9-0.691 X10+0.914 X11-3.943 X12-1530.478=1496.878
Second discrimination function FON=7.556 X1+4.831 X2+8.999 X3+102.984 X4+2.680 X5-0.960X6-2.482 X7+3.491 X8+0.857 X9-0.690 X10+1.735 X11-4.203 X12-1550.198=1491.544
Analyzed, determined and result output module compared FOCValue sum FONValue, FOC>FONAnd outputting a prediction result that the probability of the ovarian malignant tumor of the detected object is more than or equal to 88.9 percent.
The tested person is diagnosed with ovarian cancer, and the clinical diagnosis result of the tested person is consistent with the prediction result of the kit provided by the invention.
Example 6 Using the System of example 1 and the kit of example 2 to predict the risk of ovarian malignancy in a subject
The detected object is: women, age 45, visiting the department of obstetrics and gynecology of the second hospital of Jilin university, performing laparotomy, with sufficient informed examination purpose and use, under the premise of their own wishes, signed an informed consent, and collected 1mL of anticoagulated blood via the peripheral vein.
The same treatments and tests were carried out on blood samples, with reference to the prediction method of example 3, and the results show that: l is1=270.94,L2=270.94,L3=403.13,L4=403.13,L5=228.65,L6=228.65,L7=400.46,L8=402.48,L9=318.18,L10=322.50,L11=259.32,L12=262.94。
Calculated according to the fragment length and the following formula, denoted X1-X12Where round stands for rounded integer:
X1=round[(L1-191)/3]=27;X2=round[(L2-191)/3]=27;
X3=round(L3-379)=24;X4=round(L4-379)=24;
X5=round[(L5-202)/2]=13;X6=round[(L6-202)/2]=13;
X7=round[(L7-359)/2]=21;X8=round[(L8-359)/2]=22;
X9=round[(L9-278)/2]=20;X10=round[(L10-278)/2]=22;
X11=round[(L11-200)/4]=15;X12=round[(L12-200)/4]=16。
using a computer running the system for predicting susceptibility to ovarian malignancy described in example 1, a susceptibility to developing ovarian malignancy in a subject is predicted by:
inputting the age, the sex and the STR locus short tandem sequence repetition times of the detected object into a system through a data input module, and calculating the result of a discriminant function through a data calculation module:
first discriminant function FOC=7.213 X1+5.611 X2+8.722 X3+102.277 X4+3.013 X5-1.076X6-3.097 X7+3.897 X8+0.695 X9-0.691 X10+0.914 X11-3.943 X12-1530.478=1474.944
Second discrimination function FON=7.556 X1+4.831 X2+8.999 X3+102.984 X4+2.680 X5-0.960X6-2.482 X7+3.491 X8+0.857 X9-0.690 X10+1.735 X11-4.203 X12-1550.198=1479.62
Analyzed, determined and result output module compared FOCValue sum FONValue, FOC≤FONAnd outputting a prediction result that the probability that the detected object does not have ovarian malignant tumor is more than or equal to 83.3 percent.
The tested object is diagnosed as teratoma after the visit, belongs to benign tumor, and the clinical diagnosis result of the tested object is consistent with the prediction result of the kit.
The foregoing is a preferred embodiment of the present invention and is not intended to limit the present invention, and it should be understood that any changes, modifications, substitutions and alterations (e.g., addition, subtraction, change of STR sites, use of cells or tissues from other sources, use of other statistical methods, etc.) made without departing from the principles and spirit of the present invention are intended to be included within the scope of the present invention.
SEQUENCE LISTING
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Claims (10)

1. A kit for predicting susceptibility to ovarian malignancy, comprising the following components: STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer and STR-6 primer, wherein the STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer and STR-6 primer are respectively used for amplifying target fragments containing the following short tandem sequences so as to determine the repetition times of the following short tandem sequences:
2. the kit for predicting susceptibility to ovarian malignant tumors is characterized by comprising the following components: STR-1 primer, STR-2 primer, STR-3 primer, STR-4 primer, STR-5 primer and STR-6 primer;
the sequences of the STR-1 primer, the STR-2 primer, the STR-3 primer, the STR-4 primer, the STR-5 primer and the STR-6 primer are as follows:
preferably, the STR-1 primer, the STR-2 primer, the STR-3 primer, the STR-4 primer, the STR-5 primer and the STR-6 primer are all used at the concentration of 10 mu M.
3. The kit of claim 2, further comprising: PCR amplification reaction liquid, LIZ-500 molecular weight internal standard and deionized formamide.
4. The kit of claim 3, wherein: the PCR amplification reaction solution is a mixed solution of the following reagents: TaqDNA polymerase 5U/. mu. L, Tris-HCl 100mM, KCl 500mM, ethylphenylpolyethylene glycol 0.8% (v/v), MgCl225mM, dNTP 10mM and deionized water; wherein Tris-HCl has a pH of 8.8 at 25 ℃.
5. The kit of claim 4 for predicting susceptibility to ovarian malignancy, wherein: further comprising instructions for use;
the application instruction records a use method of the ovarian malignant tumor susceptibility prediction kit, which comprises the following steps:
(1) extracting sample DNA;
(2) PCR reaction
(2-1) taking out the STR-1 primer, the STR-2 primer, the STR-3 primer, the STR-4 primer, the STR-5 primer, the STR-6 primer and the PCR amplification reaction solution from a refrigerator, balancing to room temperature, fully dissolving each component, and respectively and rapidly centrifuging for 10 seconds;
(2-2) adding 30-300ng of sample DNA into 60 mu L of PCR amplification reaction solution, adding deionized water to supplement to 115.2 mu L, fully and uniformly mixing, quickly centrifuging for 10 seconds, and subpackaging the mixed solution into 6 PCR reaction tubes according to 19.6 mu L/hole;
(2-3) respectively adding an STR-1 primer, an STR-2 primer, an STR-3 primer, an STR-4 primer, an STR-5 primer and an STR-6 primer into the 6 PCR reaction tubes in the step (2-2) according to 0.8 mu L/hole; covering a PCR reaction tube cover, recording the sample adding condition, quickly centrifuging for 10 seconds, then transferring the PCR reaction tube to a corresponding position of a sample groove of a PCR amplification instrument, recording the placing sequence, and starting the PCR amplification reaction; the amplification reaction conditions are as follows: 3 minutes at 95 ℃; 30 seconds at 95 ℃, 30 seconds at 60 ℃ and 30 seconds at 72 ℃ for 10 cycles; 30 seconds at 95 ℃, 30 seconds at 55 ℃, 30 seconds at 72 ℃ and 20 cycles; 6 groups of PCR amplification products are obtained at 72 ℃ for 6 minutes;
(3) STR fragment analysis
(3-1) adding 990 mu L of deionized formamide into 10 mu L of LIZ-500 molecular weight internal standard, fully and uniformly mixing, quickly centrifuging for 10 seconds, respectively adding into a sequencing reaction tube according to 10 mu L/hole, and quickly centrifuging for 10 seconds;
(3-2) adding the 6 groups of PCR amplification products into 6 sequencing reaction tubes according to 1 mu L/hole respectively, and quickly centrifuging for 10 seconds; then transferring the sequencing reaction tube to a corresponding position of a sample tank of a PCR (polymerase chain reaction) amplification instrument, heating at 98 ℃ for 5 minutes, immediately placing the sequencing reaction tube on an ice-water mixture after the program is finished, rapidly cooling to 0 ℃, and rapidly centrifuging for 10 seconds; then transferring the sequencing reaction tube to a corresponding position of a sample groove of an STR locus fragment analyzer, recording the placement sequence, and performing fragment analysis detection;
(4) analysis and determination of results
(4-1) respectively recording the fragment lengths of two alleles at each site of STR-1, STR-2, STR-3, STR-4, STR-5 and STR-6 according to the fragment analysis result:
the length of the smaller of the two STR-1 alleles is recorded as L1And the length of the larger fragment of the two STR-1 alleles is designated as L2
The length of the smaller fragment of the two STR-2 alleles is recorded as L3And the length of the larger fragment of the two STR-2 alleles is designated as L4
The length of the smaller fragment of the two STR-3 alleles is recorded as L5And the length of the larger fragment of the two STR-3 alleles is marked as L6
The length of the smaller fragment of the two STR-4 alleles is recorded as L7And the length of the larger fragment of the two STR-4 alleles is marked as L8
The length of the smaller of the two STR-5 alleles is recorded as L9And the length of the larger fragment of the two STR-5 alleles is designated as L10
The length of the smaller fragment of the two STR-6 alleles was designated L11And the length of the larger fragment of the two STR-6 alleles is marked as L12
(4-2) the number of repetitions of the short tandem sequence is calculated from the fragment length and the following formula, and is denoted as X1-X12Where round stands for rounded integer:
X1=round[(L1-191)/3];X2=round[(L2-191)/3];
X3=round(L3-379);X4=round(L4-379);
X5=round[(L5-202)/2];X6=round[(L6-202)/2];
X7=round[(L7-359)/2];X8=round[(L8-359)/2];
X9=round[(L9-278)/2];X10=round[(L10-278)/2];
X11=round[(L11-200)/4];X12=round[(L12-200)/4];
(4-3) substituting the number of the short tandem sequence repetitions into a preset discriminant function:
FOC=7.213X1+5.611X2+8.722X3+102.277X4+3.013X5-1.076X6-3.097X7+3.897X8+0.695X9-0.691X10+0.914X11-3.943X12-1530.478
FON=7.556X1+4.831X2+8.999X3+102.984X4+2.680X5-0.960X6-2.482X7+3.491X8+0.857X9-0.690X10+1.735X11-4.203X12-1550.198;
(4-4) prediction of susceptibility to ovarian malignancy:
comparison FOCValue sum FONValue if FOC>FONPredicting the probability of the tested object suffering from ovarian malignant tumor to be more than or equal to 88.9 percent; if FOC≤FONAnd predicting that the probability that the detected object does not suffer from the ovarian malignant tumor is more than or equal to 83.3 percent.
6. The kit of claim 5, wherein: the sample is whole blood of a subject.
7. Use of the ovarian malignancy susceptibility pre-test kit according to any one of claims 1 to 6 in the preparation of an ovarian malignancy diagnostic product.
8. A ovarian malignancy susceptibility prediction system, comprising:
means for obtaining the number of repetitions of the following short tandem STR loci of the sample DNA:
data processing and decision device, comprising the following modules:
the data input module is used for inputting the age, the sex and the STR locus short tandem sequence repetition times of the detected object;
the database management module is used for the operation management of data storage, modification, deletion, inquiry and printing;
the data calculation module is used for calculating a discrimination function result according to the repeat times of the STR locus short serial sequence in the data input module;
and the analysis, discrimination and result output module is used for comparing the discrimination function results so as to predict the susceptibility of the ovarian malignant tumor and output the result.
9. The system of claim 8, wherein the number of times X of the STR site short tandem repeat of the sample DNA obtained by the kit of claim 2 is used to predict susceptibility to ovarian malignancy1-X12(ii) a And
wherein:
the discriminant function includes:
first discriminant function FOC=7.213X1+5.611X2+8.722X3+102.277X4+3.013X5-1.076X6-3.097X7+3.897X8+0.695X9-0.691X10+0.914X11-3.943X12-1530.478
Second discrimination function FON=7.556X1+4.831X2+8.999X3+102.984X4+2.680X5-0.960X6-2.482X7+3.491X8+0.857X9-0.690X10+1.735X11-4.203X12-1550.198
In the case of the discriminant function,
X1the number of repeats of the short tandem sequence for the smaller of the two alleles of STR-1;
X2the number of repeats of the short tandem sequence for the larger of the two alleles of STR-1;
X3the number of repeats of the short tandem sequence for the smaller of the two alleles of STR-2;
X4the number of repeats of the short tandem sequence for the larger of the two alleles of STR-2;
X5two for STR-3The number of repeats of the short tandem sequence of the smaller fragment in the allele;
X6the number of repeats of the short tandem sequence for the larger of the two alleles of STR-3;
X7the number of repeats of the short tandem sequence for the smaller of the two alleles of STR-4;
X8the number of repeats of the short tandem sequence for the larger of the two alleles of STR-4;
X9the number of repeats of the short tandem sequence for the smaller of the two alleles of STR-5;
X10the number of repeats of the short tandem sequence for the larger of the two alleles of STR-5;
X11the number of repeats of the short tandem sequence for the smaller of the two alleles of STR-6;
X12the number of repeats of the short tandem sequence for the larger of the two alleles of STR-6;
wherein, X1 -X12Calculated from the fragment length and the following, where round stands for rounded integer:
X1=round[(L1-191)/3];X2=round[(L2-191)/3];
X3=round(L3-379);X4=round(L4-379);
X5=round[(L5-202)/2];X6=round[(L6-202)/2];
X7=round[(L7-359)/2];X8=round[(L8-359)/2];
X9=round[(L9-278)/2];X10=round[(L10-278)/2];
X11=round[(L11-200)/4];X12=round[(L12-200)/4];
X1 -X12in, L1Is the smaller segment length value, L, of the two alleles of STR-12Is in two alleles of STR-1A larger segment length value;
L3is the smaller segment length value, L, of the two alleles of STR-24Is the larger fragment length value of the two alleles of STR-2;
L5is the smaller segment length value, L, of the two alleles of STR-36Is the larger fragment length value of the two alleles of STR-3;
L7is the smaller segment length value, L, of the two alleles of STR-48Is the larger fragment length value of the two alleles of STR-4;
L9is the smaller segment length value, L, of the two alleles of STR-510Is the larger fragment length value of the two alleles of STR-5;
L11is the smaller segment length value, L, of the two STR-6 alleles12Is the larger fragment length value of the two alleles of STR-6;
the analysis discrimination and result output module outputs a first discrimination function FOCAnd a second discrimination function FONIf F is the result of the calculation ofOC>FONOutputting a prediction result that the probability of the ovarian malignant tumor of the detected object is more than or equal to 88.9%; if FOC≤FONAnd outputting a prediction result that the probability that the detected object does not suffer from ovarian malignant tumor is more than or equal to 83.3 percent.
10. Use of the system of claim 8 in the preparation of ovarian malignancy prediction products, ovarian malignancy diagnostic products, and ovarian health-aid products.
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