阅读说明：本技术 一种用于跨物种个体识别方法及个体识别分析系统 (Cross-species individual identification method and individual identification analysis system ) 是由 李梦瑶 赵梓丞 贺小兰 姜鑫然 陈银 王轶男 于 2021-09-17 设计创作，主要内容包括：本发明属于遗传学测定技术领域,尤其为一种用于跨物种个体识别方法及个体识别分析系统,包括以下步骤：步骤一、获取STR基因座组；101)、将包括人、猪、牛、马、山羊、狗、猫、绵羊、兔子、牦牛在内的十种常见物种的全基因序列中存在核心基因序列取出；本发明创造技术相比于现有技术方案最重要的优点就是相较于现有单一物种的个体识别和亲缘鉴定实现了对常见的十种物种种类跨物种鉴别,使他们共用一种STR位点,极大地简化了单一物种各自维护一个对应的DNA分析系统的相关工作,对应多个物种只需共用一个DNA分析系统即可,利用本发明提供的STR基因座集与个体识别算法可以实现以上十种物种的个体识别和亲缘鉴定。(The invention belongs to the technical field of genetic determination, and particularly relates to a cross-species individual identification method and an individual identification analysis system, which comprise the following steps: step one, obtaining an STR (short tandem repeat) gene seat group; 101) taking out core gene sequences existing in the whole gene sequences of ten common species including human, pig, cattle, horse, goat, dog, cat, sheep, rabbit and yak; compared with the existing single species individual identification and relative identification, the most important advantage of the invention is that the cross-species identification of the ten species is realized, so that the species share one STR locus, the related work of maintaining one corresponding DNA analysis system for each species is greatly simplified, only one DNA analysis system is needed for sharing a plurality of species, and the individual identification and relative identification of the ten species can be realized by utilizing the STR locus set and the individual identification algorithm provided by the invention.)

1. A method for cross-species individual identification, comprising the steps of:

s1: obtaining an STR gene seat group;

s2: obtaining species DNA;

s3: and obtaining a DNA recognition result.

2. The method of claim 1, wherein in the step of S1, obtaining STR loci comprises the steps of:

s101: taking out core gene sequences existing in the whole gene sequences of ten common species including human, pig, cattle, horse, goat, dog, cat, sheep, rabbit and yak;

s102: and storing the extracted core gene sequence in a storage device to obtain an STR locus group.

3. The method for cross-species individual identification according to claim 1, wherein in the S2, species DNA is obtained, comprising the steps of:

s201: placing the blood of the species in a reaction tube;

s202: pouring cell lysis solution into a reaction test tube to lyse cells;

s203: magnetic silica gel particles are put into a reaction test tube to adsorb DNA molecules which are dissociated from the lysed cells;

s204: taking out the particles;

s205: the adsorbed DNA molecules were eluted using pure water.

4. The method for cross-species individual identification according to claim 1, wherein in the S3, obtaining DNA identification result comprises the following steps:

s301: placing the obtained DNA molecules in an individual recognition analysis system;

s302: DNA recognition results were obtained by STR locus sets using an individual recognition system.

5. The method according to claim 1, wherein the operation of S2 is performed by stirring slowly and uniformly with a glass rod for 10min during cell lysis in the reaction tube.

6. The method according to claim 1, wherein in the step of S2, when washing the DNA molecules, 70% ethanol is poured into a reaction tube containing magnetic silica gel, the reaction tube is shaken at room temperature for 10min, the clear solution on the surface of the magnetic silica gel is separated by centrifugal shaking, the magnetic silica gel is taken out, after cleaning is performed twice, pure water is added, vortex mixing is performed, and the DNA molecules are separated for 10 min.

7. An individual identification analysis system, comprising:

a data acquisition module (1), the data acquisition module (1) being for acquiring DNA molecules;

a molecular structure generation module (2), the molecular structure generation module (2) acquiring DNA molecular structure data by using a corresponding apparatus;

the characteristic extraction module (3) is used for extracting core molecular data in the molecular structure data generated by the molecular structure generation module (2), and the characteristic extraction module (3) is electrically connected with the molecular structure generation module (2);

the sequencing module (4), the sequencing module (4) is used for analyzing the data segment that the characteristic extraction module (3) sent to obtain the direction and the structure of DNA, fix a position the sudden change simultaneously, the sequencing module (4) and characteristic extraction module (3) electric connection.

8. The individual identification and analysis system of claim 7, further comprising:

the comparison module (5) carries out corresponding retrieval and comparison work according to the information sent by the receiving and sequencing module (4) and the data in the STR gene seat group to obtain an analysis result, and the comparison module (5) is electrically connected with the sequencing module (4);

the output module (6), the said output module (6) is used for revealing the analysis result that the comparison module (5) transmits, the said output module (6) is electrically connected with comparison module (5);

the information database (7), the information database (7) is used for storing STR locus group data, and the information database (7) is electrically connected with the comparison module (5);

the data updating module (8) is used for updating STR (short tandem repeat) locus group data in the information database (7), and the data updating module (8) is electrically connected with the information database (7).

Technical Field

The invention belongs to the technical field of genetic determination, and particularly relates to a cross-species individual identification method and an individual identification analysis system.

Background

A gene is a DNA or RNA sequence that carries genetic information, and a gene is a DNA or RNA fragment having a genetic effect, i.e., a genetic element. The gene is a basic genetic unit for controlling biological traits, expresses genetic information carried by the gene by guiding protein synthesis, and controls the trait expression of biological individuals. Genetic testing is a technique for detecting DNA using biological samples such as blood, other body fluids, tissues, and cells. The gene detection can be used for diagnosing genetic diseases, genetic counseling analysis, the probability of genetic diseases of family members of the genetic diseases, determining carriers of pathogenic genes and prenatal diagnosis, can also be used for predicting the risk of certain diseases such as tumors, and can also be used for paternity test and developmental identification. Gene detection is an important means for precise medical treatment. The currently common gene detection methods are: first-generation and second-generation DNA sequencing, multiple connection probe amplification, fluorescent quantitative PCR, gene chip, fluorescent in situ hybridization and other detection technologies.

The existing genetic identification or individual identification technologies are all single species identification which is implemented for single species, such as identification of single species implemented for species like human, dog, panda, etc., and the identification of each single species requires the use of a corresponding analysis system and analysis work, but the work related to maintaining a corresponding DNA analysis system for each species is expensive, time-consuming and labor-consuming.

Disclosure of Invention

In order to solve the problems in the prior art, the invention provides a cross-species individual identification method and an individual identification analysis system, which comprise a set of unified STR locus set comprising ten common species including human, pig, cow, horse, goat, dog, cat, sheep, rabbit and yak, thereby realizing cross-species identification.

In order to achieve the purpose, the invention provides the following technical scheme: a method for cross-species individual identification, comprising the steps of:

s1: obtaining an STR gene seat group;

s2: obtaining species DNA;

s3: and obtaining a DNA recognition result.

The invention is further configured to: in S1, the method for obtaining the STR locus set includes the following steps:

s101: taking out core gene sequences existing in the whole gene sequences of ten common species including human, pig, cattle, horse, goat, dog, cat, sheep, rabbit and yak;

s102: and storing the extracted core gene sequence in a storage device to obtain an STR locus group.

The invention is further configured to: in said S2, obtaining species DNA, comprising the steps of:

s201: placing the blood of the species in a reaction tube;

s202: pouring cell lysis solution into a reaction test tube to lyse cells;

s203: magnetic silica gel particles are put into a reaction test tube to adsorb DNA molecules which are dissociated from the lysed cells;

s204: taking out the particles;

s205: the adsorbed DNA molecules were eluted using pure water.

The invention is further configured to: in the S3, obtaining a DNA recognition result, comprising the steps of:

s301: placing the obtained DNA molecules in an individual recognition analysis system;

s302: DNA recognition results were obtained by STR locus sets using an individual recognition system.

The invention is further configured to: in the operation procedure according to S2, when the cells were lysed in the reaction tube, stirring work was slowly and uniformly performed using a glass rod for 10 min.

The invention is further configured to: according to the operation of S2, when washing DNA molecules, 70% ethanol is poured into a reaction tube in which magnetic silica gel is located, and the reaction tube is shaken at room temperature for 10min, and then the clear solution on the surface of the magnetic silica gel is separated by centrifugal shaking, the magnetic silica gel is taken out, and after two times of cleaning, pure water is added, and the mixture is vortexed and mixed uniformly for 10min to separate the DNA molecules.

An individual identification analysis system comprising:

a data acquisition module for acquiring DNA molecules;

a molecular structure generation module that acquires DNA molecular structure data by using a corresponding apparatus;

the characteristic extraction module is used for extracting core molecular data in the molecular structure data generated by the molecular structure generation module and is electrically connected with the molecular structure generation module;

and the sequencing module is used for analyzing the data fragments sent by the feature extraction module, acquiring the direction and the structure of the DNA, and positioning mutation, and is electrically connected with the feature extraction module.

The invention is further configured to: further comprising:

the comparison module carries out corresponding retrieval and comparison work according to the information sent by the receiving and sequencing module and the data in the STR gene seat group to obtain an analysis result, and the comparison module is electrically connected with the sequencing module;

the output module is used for displaying the analysis result transmitted by the comparison module and is electrically connected with the comparison module;

the information database is used for storing STR (short tandem repeat) locus group data and is electrically connected with the comparison module;

and the data updating module is used for updating the STR locus group data in the information database and is electrically connected with the information database.

Compared with the prior art, the invention has the beneficial effects that:

1. compared with the existing single species individual identification and relative identification, the most important advantage of the invention is that the cross-species identification of the ten species is realized, so that the species share one STR locus, the related work of maintaining one corresponding DNA analysis system for each species is greatly simplified, only one DNA analysis system is needed for sharing a plurality of species, and the individual identification and relative identification of the ten species can be realized by utilizing the STR locus set and the individual identification algorithm provided by the invention.

2. Placing a sample on a data acquisition module, simultaneously using a molecular structure generation module to carry out DNA molecular structure data memorability identification on a DNA sample on the data acquisition module and generate data information, reducing the size of DNA molecular structure data through a characteristic extraction module, improving the identification efficiency while reducing the identification burden of a system, then using a sequencing module to calculate according to DNA molecular core structure data and search rules to obtain the direction and the structure of the DNA, carrying out positioning recording on a mutation position and sending the recorded characteristics to a comparison module, retrieving STR (short tandem repeat) locus group information recorded in an information database, obtaining a final detection result according to the STR locus information, identifying STR loci of species DNA, carrying out individual identification by comparing whether the STR loci corresponding to the front and the back of an animal are the same or not, and finally sending the individual identification result to an output module for display work, the use of STR loci as a whole allows for individual identification and genetic identification of the ten species mentioned above, for which STR loci have high individual identification capability through the application of individual identification algorithms.

Drawings

The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention and not to limit the invention. In the drawings:

FIG. 1 is a flow chart of an identification method of the present invention;

FIG. 2 is a system diagram of an individual identification analysis system according to the present invention.

In the figure: 1. a data acquisition module; 2. a molecular structure generation module; 3. a feature extraction module; 4. a sequencing module; 5. a comparison module; 6. an output module; 7. an information database; 8. and a data updating module.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Referring to fig. 1, the present invention provides the following technical solutions: a method for cross-species individual identification, comprising the steps of:

step one, obtaining an STR (short tandem repeat) gene seat group;

101) taking out core gene sequences existing in the whole gene sequences of ten common species including human, pig, cattle, horse, goat, dog, cat, sheep, rabbit and yak;

102) storing the extracted core gene sequence in a storage device to obtain an STR (short tandem repeat) locus group;

step two, obtaining species DNA;

201) placing the blood of the species in a reaction tube;

202) pouring cell lysis solution into the reaction tube to lyse cells, and slowly and uniformly stirring the cells in the reaction tube for 10min by using a glass rod when the cells are lysed;

203) putting magnetic silica gel particles into a reaction test tube to adsorb DNA molecules which are dissociated from the lysed cells;

204) taking out the particles;

205) eluting the adsorbed DNA molecules by using pure water, pouring 70% ethanol into a reaction tube in which the magnetic silica gel is positioned when washing the DNA molecules, shaking for 10min at room temperature, separating clear liquid on the surface of the magnetic silica gel by centrifugal shaking, taking out the magnetic silica gel, repeatedly cleaning for two times, adding the pure water, uniformly mixing in a vortex manner, and separating the DNA molecules for 10 min;

step three, obtaining a DNA recognition result;

301) placing the obtained DNA molecules in an individual recognition analysis system;

302) and obtaining a DNA recognition result through the STR gene locus group by using the individual recognition system.

According to the invention, the core gene sequence data obtained from the whole gene sequences of various species are stored in the storage device, so that the space occupied by the gene sequence data can be reduced, and meanwhile, the various gene sequences are stored together, so that the uniform STR analysis of various species is realized, and the cost consumed by the analysis result is reduced by using the same analysis system;

when an STR locus set is obtained, a special reference system is adopted, because the STR locus set provided by the invention comprises ten different species, and chromosomes of each species are different, in order to solve the problem, the special reference system is created, and the specific implementation method is as follows. First, we aligned the original sequencing data to a reference for the human genome, processed the data through the lobSTR workflow, and obtained STR locus candidate sets from overlapping genomic regions of related species. We exclude loci and single nucleotide repeats on sex chromosomes as these are not applicable in practical situations. Using the loci retrieved, we estimated the allele frequencies of each locus and used these frequencies to calculate PD, PM, PE, HE and the maximum genotype frequencies. Then we apply the threshold values specified in the equation (with the following requirements for PD ≧ delta for different sites in each species respectively_d、PM≤δ_m、PE≥δ_e、HE≥δ_h；(δ_d＝0.6，δ_m＝0.4，δ_e＝0.3，δ_h0.5) STR loci were filtered on PD, PM, PE, HE, and call rate. The method is a realization process of a special reference system, a group of unique databases can be obtained from the reference system and used as standard reference systems of ten species, and gene loci corresponding to the gene loci in the database can be obtained through the backbone so as to realize the unification of the cross-species chromosome gene loci;

when species DNA is obtained, the cell lysis solution is used for lysing cells, the magnetic silica gel particles are used for extracting DNA molecules in a sample in a matching way, a plurality of reaction tubes are avoided being used during extraction, so that the integrity of DNA molecule sample extraction is improved, the whole operation process is simpler, finally, when the DNA sample comparison work is carried out, a related individual identification system can be avoided being used, compared with the existing individual identification and affinity identification of a single species, the cross-species identification of ten species is realized, so that the species share one STR locus, the related work of maintaining a corresponding DNA analysis system by each single species is greatly simplified, only one DNA analysis system is needed to share corresponding to a plurality of species, meanwhile, a large number of experiments are carried out on the invention, and the experimental results show that, for single species and mixed populations, the unified set of loci has a high integrated discriminative power, >1-10^ -9, and random match probabilities for all species involved, <10^ -7, indicating that the identified set of STR loci can be applied to multiple species. In the industry, STR sites whose general forensic parameters meet the following threshold requirements can be used as forensic identification bases, including: HE (heterozygosity) > 0.6, PD (individual discrimination ability) > 0.6, PIC (polymorphic locus utility value) > 0.5, and CPD (cumulative individual discrimination ability) > 0.99999. Corresponding to the STR locus provided by the invention, the experimental result shows that CPD, PD and CPI of the STR locus corresponding to the invention all meet the relevant standards, and the STR locus set and the individual identification algorithm provided by the invention can realize the individual identification and the relative identification of the ten animals.

As shown in fig. 2, an individual recognition analysis system includes:

the data acquisition module 1, the data acquisition module 1 is used for obtaining DNA molecules;

a molecular structure generation module 2, the molecular structure generation module 2 acquiring DNA molecular structure data by using a corresponding apparatus;

the characteristic extraction module 3 is used for extracting core molecule data in the molecular structure data generated by the molecular structure generation module 2, and the characteristic extraction module 3 is electrically connected with the molecular structure generation module 2;

and the sequencing module 4 is used for analyzing the data fragments sent by the feature extraction module 3, acquiring the direction and the structure of the DNA, positioning the mutation, and electrically connecting the sequencing module 4 with the feature extraction module 3.

The invention is further configured as an individual identification analysis system, further comprising:

the comparison module 5 is used for carrying out corresponding retrieval and comparison work according to the information sent by the receiving and sequencing module 4 and the data in the STR gene seat group to obtain an analysis result, and the comparison module 5 is electrically connected with the sequencing module 4;

the output module 6 is used for displaying the analysis result transmitted by the comparison module 5, and the output module 6 is electrically connected with the comparison module 5;

the information database 7 is used for storing STR (short tandem repeat) locus group data, and the information database 7 is electrically connected with the comparison module 5;

and the data updating module 8 is used for updating the STR locus group data in the information database 7, and the data updating module 8 is electrically connected with the information database 7.

When the invention works, a sample is placed on a data acquisition module 1, a molecular structure generation module 2 is used at the same time to carry out DNA molecular structure data memorability identification on a DNA sample on the data acquisition module 1, generate data information and send the data information to a feature extraction module 3, the feature extraction module 3 carries out extraction work on core structure data in the DNA molecular structure data, the size of the DNA molecular structure data is reduced, the identification load of a system is reduced, the identification efficiency can be improved, then the DNA molecular core structure data is sent to a sequencing module 4, the sequencing module 4 carries out calculation according to the DNA molecular core structure data, a rule is searched to obtain the direction and the structure of DNA, the mutation position is positioned and recorded, the recorded feature is sent to a comparison module 5, the comparison module 5 searches STR gene seat group information recorded in an information database 7 according to the received feature information, and obtain the final test result according to STR gene group information, discern STR locus of species DNA, carry on the individual identification by comparing the corresponding STR locus before and after an animal is the same, send the individual identification result to the output module 6 to carry on the display work finally, can realize the individual identification and the relative identification to the above-mentioned ten kinds of species with STR gene group set on the whole use, have high individual identification ability to these STR loci through using the individual identification algorithm, when needing to revise STR gene group data, can use the data updating module 8, revise the data in the information database 7, improve the expanding use effect of the system.

Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.