CN106906300A - A kind of genetic ID card and preparation method thereof - Google Patents
A kind of genetic ID card and preparation method thereof Download PDFInfo
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- CN106906300A CN106906300A CN201710279234.8A CN201710279234A CN106906300A CN 106906300 A CN106906300 A CN 106906300A CN 201710279234 A CN201710279234 A CN 201710279234A CN 106906300 A CN106906300 A CN 106906300A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
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- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
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Abstract
The present invention relates to DNA or gene applied technical field, in particular to a kind of genetic ID card and preparation method thereof, the genetic ID card includes：Storage medium；The gene identities number of the specific gene fragment sequence of individual distinctive 16 locus of reflection are recorded on the storage medium., also there is substantial amounts of STR bit point in the site of the existing genetic marker of locus on genetic ID card, therefore the site of gene pleiomorphism has generality, and discrimination is high, it may also be used for paternity test.When genetic ID card is made, 16 information of locus are also beneficial to the prediction and diagnosis for carrying out disease.
Description
Technical field
The present invention relates to DNA or gene applied technical field, in particular to a kind of genetic ID card and its preparation side
Method.
Background technology
Now widely used identity card, only carrier surface describe card owner general essential information, such as name,
Graphical information in terms of the text information and portrait of the aspects such as sex, native place, date of birth, though some social concerns can be solved.
But, can change due to them and forge, often it is difficult to judge the owner that holder is exactly real card, learned in crime field and led
Domain is particularly true.The country such as American and Britain has set up various DNA fingerprint databases, but can only solve asking for some criminology fields
Topic, and more individuals, family, medical treatment and social concern can not be solved.
Genetic ID card (Gene Identification Card) mainly uses DNA fingerprint technology, chooses several
Fixed gene loci is identified.Gene is intrinsic constant genetic marker, is inherited from father and mother there by children, only
Several sites need to be chosen and can just be identified.Genetic ID card and regular identity card are in profile and substance without many
Big difference, in ID card No., ID card No. originally is by representing the features such as region, date of birth, sex to Main Differences
Numeral composition.
But existing genetic ID card function is more single, also not high enough, the selected site of the degree of accuracy that discriminates one's identification
Only for used by authentication, and cost of manufacture is higher.
In view of this, it is special to propose the present invention.
The content of the invention
It is an object of the invention to provide a kind of genetic ID card, described genetic ID card can solve the problem that discrimination it is low,
Detection of complex, the big technical problem of number of repetition difference.
A kind of genetic ID card, including：Storage medium；
The gene of the specific gene fragment sequence of individual distinctive 16 locus of reflection is recorded on the storage medium
ID；
16 locus be respectively DPYD, CYP2D6, SULT1A1, CYP3A4, CYP2C9, CYP19A1, ITPA,
MDR1, ADH2, COX-2, MTHFR-C677T, GSTM1, GSTT1, GSTP1, PPAR- α and PPAR- γ.
Preferably, the storage medium is chip, CD or magnetic material, it is furthermore preferred that the storage medium is arranged at
On one card.
Selected 16 locus of the present invention, are the genetic fragments with numerous STR or RFLP pleomorphism sites, many
Limited loci information collectively form specific gene fingerprint so that the probability pole that is repeated with different people
Low, its human specific of software statistics is high, and discrimination is up to more than 99.9999%.
Additionally, diagnosis of the said gene seat to disease is related, the polymorphism of such as DPYD is close with the incidence of disease of the carcinoma of the rectum
Cut is closed, and the neurological susceptibility of the polymorphism of SULT1A1 then to the disease such as fibroid, the cancer of the esophagus is related.Therefore, said gene is prepared
During identity card, the gene screening of numerous diseases is also completed simultaneously, achieve many things at one stroke.
Preferably, genetic ID card as described above, states the individual essential information that is also stored with storage medium.
Preferably, genetic ID card as described above, the individual essential information includes abo blood group, Rh blood groups information, shines
Piece, name, sex, the date of birth, address, father and mother's name, ID card information, familial inheritance medical history, allergies, previous conviction and
One or more in bad habit.
Present invention also offers a kind of preparation method of said gene identity card, the method includes：
Individual specific gene fragment sequence is obtained, the storage that the specific gene fragment sequence is stored in identity card is situated between
In matter；
Preferably, methods described also using the individual finger print information or pupil information to the storage medium
In the specific gene fragment sequence be encrypted.
Specifically, the sequences of 16 genetic fragments first are surveyed into logical, then it is input into computer and is encrypted, AES can be with
The algorithm being known in the art so that specific sequence information encryption is stored in the storage medium of genetic ID card and/or takes
Business device end；
Also, it is optional, the information after encryption is corresponded with unique identification code, and the identification code is printed upon
The surface of the genetic ID card.
Preferably, described information encryption is specially and for the specific gene fragment sequence to be stored in fingerprint or pupil
In the microchip of encryption.
Preferably, method as described above, the individual specific gene fragment sequence of the acquisition comprises the following steps：
1) DNA profiling of party, is extracted；
2), through the specific gene fragment of PCR amplification target gene seats；
3), the specific gene fragment is sequenced.
Preferably, in step 2) in, when carrying out the PCR and expanding, for expand DPYD, CYP2D6, SULT1A1,
CYP3A4、CYP2C9、CYP19A1、ITPA、MDR1、ADH2、COX-2、MTHFR-C677T、GSTM1、GSTT1、GSTP1、
The sense primer of PPAR- α and PPAR- γ is successively such as SEQ ID NO：Shown in 1-16；Anti-sense primer is successively such as SEQ ID NO：
Shown in 17-32.
Composition of the present invention has specific good, amplification effect by cleverly design of primers, 16 pairs of upstream and downstream primers
Rate advantage high, will not form primer dimer, non-specific amplification and cross reaction in PCR amplification procedures, it is to avoid non-purpose
Influence of the appearance of product to genotyping result, while amplification efficiency is high, single PCR reactions can simultaneously amplify 16 sequences,
Coordinate high throughput sequencing technologies, be greatly improved the detection efficiency of amplified production.In view of genetic ID card on a large scale should following
Used time, sample size is very large, thus genetic ID card prepare when improved efficiency can save great amount of cost.
Preferably, method as described above, in step 2) in, when carrying out the PCR amplifications, by sequence such as SEQ ID NO:
Primer shown in 1-32 is expanded in being blended in same amplification system；
Preferably, in some embodiments, sequence such as SEQ ID NO:The molar concentration between primer shown in 1-32
Than being followed successively by 0.7~1.0：0.3~0.9：1.0~1.2：0.4~0.6：0.7~1.0：0.5~1.0：1.0~1.2：1.0~
1.2：1.4~1.6：0.8~1.0：1.0~1.2：1.0~1.2：1.4~1.6：1.8~2.0：1.8~2.0：1.8~2.0：
0.7~1.0：0.3~0.9：1.0~1.2：0.4~0.6：0.7~1.0：0.5~1.0：1.0~1.2：1.0~1.2：1.4~
1.6：0.8~1.0：1.0~1.2：1.0~1.2：1.4~1.6：1.8~2.0：1.8~2.0：1.8~2.0.
Preferably, in some embodiments, in step 2) in, when carrying out the PCR amplifications, annealing temperature is 60~62
DEG C, cycle-index is 28~32 times.
It is furthermore preferred that the condition of PCR reactions is：1st step, 94~95 DEG C of 3~5min of denaturation；2nd step, 94~95 DEG C of denaturation
7~10s；3rd step, 60~62 DEG C of 0.5~1min of annealing；4th step, 71~72 DEG C of 25~30s of extension；5th step, repeats the 2nd~4
Step 28~32 times；6th step, 59~60 DEG C of 9~10min of extension；7th step, 4~5 DEG C of lasting insulations.
Preferably, in some embodiments, the DNA profiling comes autoblood, saliva, seminal fluid, bone or hair.
Preferably, in some embodiments, the DNA profiling passes through saturation phenol chloroform method, resins extraction method or magnetic
Pearl extraction method is extracted, or, the DNA profiling is the blood filter paper containing DNA, saliva card or FTA cards.
Compared with prior art, beneficial effects of the present invention are：
First, the site of present invention selection is all the site of high polymorphism in crowd.This polymorphism refers to DNA polymorphic
Property, this kind of site is selected because it is most people and has, belong to conservative gene site, stability is higher, thus site
Contained information content will be larger, and the result of identification is also more reliable.
Second, the site of selection is all the disease related locus with Familial Occurrence.Site on genetic ID card was both
There is the site of genetic marker, also there is substantial amounts of STR bit point, therefore the site of gene pleiomorphism has generality, discrimination is high,
Can be additionally used in paternity test.When genetic ID card is made, 16 information of locus are also beneficial to carry out the prediction of disease and examine
It is disconnected.
3rd, 16 pairs of upstream and downstream primers have the advantages that specific good, amplification efficiency is high, and single PCR reactions can simultaneously
16 sequences are amplified, coordinate high throughput sequencing technologies, improved efficiency when prepared by genetic ID card can save great amount of cost,
The related application of backstepping is being carried out, such as during the identity authentication of unknown corpse, it is also possible to reduces cost, beneficial to genetic ID card
Promote and application.
Specific embodiment
Genetic ID card of the present invention, because recording the individual str locus type information i.e. individual with height individual specificity
Gene identities code, thus it has the proof of identification power of height, is a kind of maximally effective proof of identification.Gene identities of the invention
The purposes and its significance of card are summarized as follows：
Gene is the fragment for including a person ownership hereditary information, is had with life, and keep all the life constant.This heredity
Information is lain in all tissues such as bone, hair, the blood of people or organ.Various genetic markers have been developed in recent years
For individual identification.Wherein STR (Short Tandem Repeat, STR) is because detection method is easy, fast
The fast, degree of accuracy is high, amplified fragments are of moderate size, and the topmost individual identification detection mark in each medical jurisprudence laboratory is had developed at present
Note.
There are many STR bit points, comprehensive these positions the invention provides 16 genomic informations, and on each genome
Point information, personal individual identification rate does not have two str locus of individuality more than 1/100000000000th between i.e. 100,000,000,000 people
Type is repeated, and personal establishing identity can be carried out completely.
Meeting with an accident, scatter, the inheritance of property, test-tube baby, bone-marrow transplantation, clone organ or cloning life forms body etc.
Reason is carried out in individual identification and paternity identification the need for causing, and genetic ID card will play vital effect.
The superiority of genetic ID card is mainly manifested in human organ transplant, blood transfusion, drug resistant gene and stem cell transplantation
The aspects such as identification.When people need donor organ and bone-marrow transplantation, can be found with crt gene identity card, especially Chinese mesh
It is preceding to set up human gene bank, then doctor can be quickly found from gene pool tissue matching identical organ, blood or
Cell, patient is succoured with most fast speed.If possessing genetic ID card before marriage, more fully examined before marriage just as having done
Look into, the generation of nearly all genetic disease can be avoided, research, paternity test, blood relationship family tree in familial disease is sought
The aspects such as confirmation are looked for, genetic ID card is even more with wide application space.
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the present invention.It is unreceipted specific in embodiment
Condition person, the condition advised according to normal condition or manufacturer is carried out.Agents useful for same or the unreceipted production firm person of instrument, are
The conventional products that can be obtained by commercially available purchase.
Embodiment
First, sample treatment
The extraction of DNA：Various methods such as including ChELEX-100 methods and phenol/chloroform method；The specific steps of first two method
It is as follows：
1), ChELEX-100 methods, a, separation karyocyte；B, the ChELEX-100 suspensions and egg that add appropriate 5-10%
White enzyme K；C, 56 DEG C of digestion d, boiling in 1-2 hours are boiled 10 minutes；E, it is quenched 3 minutes on ice, -20 DEG C preserve standby inspection or continue directly to the
3 steps.
2), phenol/chloroform method a, separation karyocyte or the tissue containing DNA；The μ l of b, SDS 250, plus phenol/chloroform/isoamyl alcohol
500 μ 1 (25: 24: 1), are centrifuged 15000rpm × 10min.
C, supernatant are transferred to the phenol/chloroform/isoamyl alcohol of 500 μ l, vibration, and centrifugation 15000rpm × 5min, supernatant are transferred to
Chloroform/the isoamyl alcohol (24: 1) of 500 μ l, vibration, the 1000 μ l that centrifugation 15000rpm × 5min, supernatant are transferred to advance ice are anhydrous
Ethanol, adds 30 μ l NaCl liquid (5M), after shaking, it is seen that DNA agglomerates.
F, separately take 0.5ml conical pipes, mark, plus 500 μ l 70% ethanol, by DNA groups choose into；After vibration, centrifugation
15000/rpm×10min。
G, supernatant is removed, it is as far as possible that water is clean (preventing DNA from pouring out).
H, often pipe add 3 steaming water dissolves DNA (4 DEG C overnight) according to amount of DNA, and -20 DEG C preserve standby inspection or continue directly to the 3rd step.
Prevent outside above-mentioned CHELECX-100 methods and phenol/chloroform method, can also use salting out method, boiling process, immunomagnetic beads method
Deng.
3), the general principle that DNA is extracted：After the multi-step of mentioned reagent is acted on, make DNA molecular with it in natural shape
The protein being combined under state and other impurity are separated, and open ground is present in the aqueous solution, are that the reaction of next step creates good
Condition.
The genomic DNA concentration extracted is diluted to 0.3ng/ μ l.
2nd, primer mixing
By sequence such as SEQ ID NO：Primer shown in 1-32 is mixed, and prepares primer mixture, wherein, it is described to draw
The concentration of each primer is as shown in table 1 in thing mixture.
Sequence such as SEQ ID NO in the primer mixture of table 1：The concentration of primer shown in 1-32
3rd, the preparation of PCR reaction systems
By the μ l of PCR reaction buffers 20, the μ l of primer mixture 12, the μ l of DNA profiling 4, deionized water 14 containing archaeal dna polymerase
μ l are well mixed, and are configured to 50 μ l PCR reaction systems.
4th, PCR reactions
The PCR reaction systems that will be prepared are placed in PCR instrument and are expanded, and wherein amplification condition is as follows：
1st step, 94 DEG C are denatured 5 minutes；2nd step, 94 DEG C are denatured 10 seconds；3rd step, 60.5 DEG C are denatured 1 minute；4th step, 72
DEG C extend 30 seconds；5th step, repeats 2-4 and walks 30 times；6th step, 61 DEG C extend 10 minutes, 4 DEG C of lasting insulations.
5th, it is sequenced
High-flux sequence instrument formation sequence information on the amplified production that step 4 is obtained.
6th, the generation of genetic ID card
Abo blood group, the Rh blood groups of the 1st, the typing individuality essential information on special computer software interface, including party
Information, photo, name, sex, date of birth, address, father and mother's name, ID card information, familial inheritance medical history, allergies, criminal
Crime record and bad habit；
2nd, 16 sequence informations of locus of typing on special computer software interface, respectively DPYD, CYP2D6,
SULT1A1、CYP3A4、CYP2C9、CYP19A1、ITPA、MDR1、ADH2、COX-2、MTHFR-C677T、GSTM1、GSTT1、
GSTP1, PPAR- α and PPAR- γ.
3rd, genetic ID card is ultimately produced.Specific software is encrypted to all information of the typing, will be individual
Body essential information is encoded and is stored in chip with the sequence information of 16 locus, and the information after encryption is unique with one
Identification code correspond, and by professional color printer output with the identification code genetic ID card.Decoded information is by text
The finger print information, and/or pupil information of this password, and/or individuality are constituted.In use, needing insertion genetic ID card and taking
Double authentication is carried out with decoded information.Additionally, also can be by repeating to measure the sequence information of above-listed locus, and by these sequences
Information input solution code system recalls individual essential information.
The genetic ID card for using the above method to make is card-like, personal information is configured with thereon and records area, the differentiation
The text information such as area and personal name is recorded for individual's front portrait without a hat on record area；Personal gene identities are additionally provided with card
Code records area.
Finally it should be noted that：Various embodiments above is merely illustrative of the technical solution of the present invention, rather than its limitations；To the greatest extent
Pipe has been described in detail with reference to foregoing embodiments to the present invention, but it will be understood by those within the art that：Its
The technical scheme described in foregoing embodiments can still be modified, or to which part or all technical characteristic
Carry out equivalent；And these modifications or replacement, the essence of appropriate technical solution is departed from various embodiments of the present invention skill
The scope of art scheme.
SEQUENCE LISTING
<110>Beijing Co., Ltd of Jun Ke genes Medical Research Institute；It is north medical data science and technology (Beijing) Co., Ltd
<120>A kind of genetic ID card and preparation method thereof
<160> 32
<170> PatentIn version 3.3
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gcggacatcg agagtataa 19
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gatccggcgt ttctcaacgc t 21
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gtaccccagt ctctacgtaa 20
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cattcagatc attgaccgc 19
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Claims (10)
1. a kind of genetic ID card, it is characterised in that including：Storage medium；
The specific gene fragment sequence of individual distinctive 16 locus of reflection is recorded on the storage medium；
16 locus be respectively DPYD, CYP2D6, SULT1A1, CYP3A4, CYP2C9, CYP19A1, ITPA, MDR1,
ADH2, COX-2, MTHFR-C677T, GSTM1, GSTT1, GSTP1, PPAR- α and PPAR- γ.
2. genetic ID card as claimed in claim 1, it is characterised in that be also stored with individual basic letter on the storage medium
Breath.
3. genetic ID card as claimed in claim 2, it is characterised in that the individual essential information includes abo blood group, Rh blood
Type information, photo, name, sex, the date of birth, address, father and mother's name, ID card information, familial inheritance medical history, allergies,
One or more in previous conviction and bad habit.
4. a kind of method for preparing the genetic ID card described in any one of claims 1 to 3, it is characterised in that including：Obtain individual
The specific gene fragment sequence of body, the specific gene fragment sequence is stored in the storage medium of identity card；
Preferably, methods described also using the individual finger print information or pupil information in the storage medium
The specific gene fragment sequence is encrypted.
5. method as claimed in claim 4, it is characterised in that the individual specific gene fragment sequence of the acquisition includes following
Step：
1) DNA profiling of party, is extracted；
2), through the specific gene fragment of PCR amplification target gene seats；
3), the specific gene fragment is sequenced.
6. method as claimed in claim 5, it is characterised in that in step 2) in, when carrying out the PCR and expanding, for expanding
DPYD、CYP2D6、SULT1A1、CYP3A4、CYP2C9、CYP19A1、ITPA、MDR1、ADH2、COX-2、MTHFR-C677T、
The sense primer of GSTM1, GSTT1, GSTP1, PPAR- α and PPAR- γ is successively such as SEQ ID NO：Shown in 1-16；Draw in downstream
Thing is successively such as SEQ ID NO：Shown in 17-32.
7. method as claimed in claim 5, it is characterised in that in step 2) in, when carrying out the PCR and expanding, by sequence such as
SEQ ID NO:Primer shown in 1-32 is expanded in being blended in same amplification system；
Preferably, sequence such as SEQ ID NO:The molar concentration rate between primer shown in 1-32 is followed successively by 0.7~1.0：0.3~
0.9：1.0~1.2：0.4~0.6：0.7~1.0：0.5~1.0：1.0~1.2：1.0~1.2：1.4~1.6：0.8~1.0：
1.0~1.2：1.0~1.2：1.4~1.6：1.8~2.0：1.8~2.0：1.8~2.0：0.7~1.0：0.3~0.9：1.0~
1.2：0.4~0.6：0.7~1.0：0.5~1.0：1.0~1.2：1.0~1.2：1.4~1.6：0.8~1.0：1.0~1.2：
1.0~1.2：1.4~1.6：1.8~2.0：1.8~2.0：1.8~2.0.
8. method as claimed in claim 7, it is characterised in that in step 2) in, when carrying out the PCR and expanding, annealing temperature
It it is 60~62 DEG C, cycle-index is 28~32 times.
9. method as claimed in claim 5, it is characterised in that the DNA profiling comes autoblood, saliva, seminal fluid, bone or hair
Hair.
10. method as claimed in claim 5, it is characterised in that the DNA profiling is carried by saturation phenol chloroform method, resin
Follow the example of or magnetic bead extraction method is extracted, or, the DNA profiling is the blood filter paper containing DNA, saliva card or FTA cards.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2019241913A1 (en) * | 2018-06-19 | 2019-12-26 | 深圳华大基因科技有限公司 | Digital identification generating method, device and system and storage medium |
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| CN103198238A (en) * | 2012-01-06 | 2013-07-10 | 深圳华大基因科技有限公司 | Drug related gene type database, gene typing and drug action detection method |
| CN105925693A (en) * | 2016-05-13 | 2016-09-07 | 深圳市核子基因科技有限公司 | Genetic identity card and preparation method thereof |
| CN106520982A (en) * | 2016-12-05 | 2017-03-22 | 中国人民解放军军事医学科学院放射与辐射医学研究所 | Compound typing system used for personal identification |
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| CN103198238A (en) * | 2012-01-06 | 2013-07-10 | 深圳华大基因科技有限公司 | Drug related gene type database, gene typing and drug action detection method |
| CN105925693A (en) * | 2016-05-13 | 2016-09-07 | 深圳市核子基因科技有限公司 | Genetic identity card and preparation method thereof |
| CN106520982A (en) * | 2016-12-05 | 2017-03-22 | 中国人民解放军军事医学科学院放射与辐射医学研究所 | Compound typing system used for personal identification |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2019241913A1 (en) * | 2018-06-19 | 2019-12-26 | 深圳华大基因科技有限公司 | Digital identification generating method, device and system and storage medium |
| US11822629B2 (en) | 2018-06-19 | 2023-11-21 | Bgi Shenzhen Co., Limited | Method and apparatus for generating digital identity and storage medium |
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