CN106520982A - Compound typing system used for personal identification - Google Patents
Compound typing system used for personal identification Download PDFInfo
- Publication number
- CN106520982A CN106520982A CN201611101246.3A CN201611101246A CN106520982A CN 106520982 A CN106520982 A CN 106520982A CN 201611101246 A CN201611101246 A CN 201611101246A CN 106520982 A CN106520982 A CN 106520982A
- Authority
- CN
- China
- Prior art keywords
- genetic marker
- snp
- typing
- identity authentication
- marker combination
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- 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
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
-
- C—CHEMISTRY; METALLURGY
- 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
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Abstract
The invention aims at providing a compound typing system used for personal identification, which is used for personal identification, especially the identification for relative relations such as grandparents and grandchild, uncles and nephews, or half sibs, and has the powerful typing efficiency and wide application range. On one aspect, the invention provides a set of genetic marker assembly, and the genetic marker assembly comprises 482 SNP sites positioned on the autosome; on another aspect, the invention provides a typing kit, and the typing kit comprises a reagent for typing the 482 SNP sites in the genetic marker assembly provided by the invention; on still another aspect, the invention provides an individual genotype identity card, and the individual genotype identity card comprises a chip, the genotype information of the genetic marker sites of the individuals is stored in the chip, and the genetic marker sites refer to the 482 SNP sites in the genetic marker assembly provided by the invention.
Description
Technical field
The invention belongs to prudence identity authentication technical field, is related to a kind of compound classification system for identity authentication,
Specifically related to 482 SNP sites and its application in identity authentication.
Background technology
Prudence DNA analysis are using modern DNA analysis technology, analysis distribution of the DNA genetic markers in colony and biography
Rule is passed, it is determined that the concordance and sibship of analysis sample, so that identity authentication is carried out to sample source sample, are that investigation is broken
Case and the judicial adjudication etc. provide technical support.Its analysis object relates generally to the various body fluid of human body (speckle), tissue, hair and table
Skin exfoliative cyte etc..Technology in prudence DNA analysis for identity authentication is primarily directed to STR at present
(Short tandem repeats, STR) and single nucleotide polymorphism (Single nucleotide polymorphisms,
SNP) typing.
STR is widely present in human genome, with high polymorphism.It is specific for one individual, on chromosome
The number of repetition of the repetitive sequence of certain ad-hoc location is fixed, and for repetition of the different individualities at same position time
Number may be different, and this just constitutes the polymorphism of these repetitive sequences in crowd.Due to this repetitive sequence in human genome
It is very many, by the detection to this polymorphism, it is possible to clearly distinguish individual different from individuality.SNP is referred to as the " third generation
DNA genetic markers ", compared with the STR of " second filial generation genetic marker " is referred to as with it is completely different the characteristics of：Densely distributed, number
Amount is huge (to be considered as the SNP genetic markers for having more than 3,000,000, this has been likely to be breached human genome in human genome
The limit of polymorphic site number), frequency high, category dimorphism labelling, exist relatively stable, therefore be considered as that application prospect is best
Genetic marker.
At present, various typing systems for identity authentication or Relationship iden- tification or examination have been developed based on SNP and STR
Agent box.For example, Chinese patent application 201310125171 discloses the typing side of 20 X-SNP sites multiple colour fluorescent compound detections
Method, 20 X-SNP sites are respectively rs5916197, rs757018 for No. rs of snp database in NCBI,
Rs2071182, rs5951622, rs4898214, rs5963947, rs5906341, rs4826645, rs3860291,
Rs5912774, rs1988916, rs5941046, rs4463614, rs5916844, rs6568051, rs2522169,
Rs5930646, rs5929739, rs5908051, rs6627351.Chinese patent application 201010265870 discloses a kind of use
In 44 SNPs sites multiple colour fluorescent multiplexed detection reagents boxes and detection method of individual identification, 44 SNP gene locis exist
In NCBI, No. rs of snp database is respectively rs1109037, rs3780962, rs987640, rs9951171, rs430046,
Rs338882, rs2342747, rs10092491, rs1821380, rs321198, rs7041158, rs13218440,
Rs560681, rs445251, rs8078417, rs10488710, rs7520386, rs214955, rs4530059,
Rs13182883, rs722290, rs6811238, rs279844, rs9905977, rs6955448, rs4288409,
Rs315791, rs740598, rs2272998, rs7205345, rs10773760, rs221956, rs576261, rs1498553,
Rs2399332, rs1058083, rs993934, rs1336071, rs1294331, rs1523537, rs2269355,
Rs12997453, rs1736442 and rs10776839.Again for example, J.M.Butler and B.Budowle et al. are disclosed and are used
19 traditional str locus seats carry out typing and identity authentication (J.M.Butler, R.Schoske, P.M.Vallone,
J.W.Redman,M.C.Kline,Allele frequencies for 15autosomal STR loci on
U.S.Caucasian African American,and Hispanic populations,J.Forensic Sci.48
(2003)908–911；J.Ge,B.Budowle,R.Chakraborty,Choosing relatives for DNA
identification of missing persons,J.Forensic Sci.56(2011)；B.Budowle,
F.R.Bieber,A.J.Eisenberg,Forensic aspects of mass disasters:strategic
Considerations for DNA-based human identification, Legal Med.7 (2005) 230 243),
This 19 str locus seats be CSF1PO, D12S391, D13S317, D16S539, D18S51, D19S433, D21S11,
D2S1338, D3S1358, D5S818, D6S1043, D7S820, D8S1179, FGA, Penta D, Penta E, THO1, TPOX with
And vWA.
Although having been developed for numerous genetic marker typing systems for identity authentication and individual identification at present, but for
Blood lineage's some sibships farther out, the compound typing system of existing genetic marker cannot make gratifying family identity mirror
Determine conclusion.For example, in practical application, existing typing system is divided to sibships such as some grandparents and grandchild, uncle and nephew or half sib
Type identification result is unsatisfactory, and the identification of such sibship is also the normal problems faceds of Jing in practical application.Therefore, for
The identification system or technology of such sibship has considerable degree of being actually needed.
In sum, it is prudence identity to develop the more powerful compound classification system of a set of typing efficiency and range of application
Major issue urgently to be resolved hurrily in identification, individual identification and sibship judgement field.
The content of the invention
The technical problem to be solved in the present invention is to provide a set of for identity authentication, is particularly suited for such as grandparent and grandchild, uncle
The Relationship iden- tification such as nephew or half sib, possess the compound classification system of more powerful typing efficiency and range of application.
On one side, the invention provides one group of genetic marker is combined, described genetic marker combination includes following 482
At least 214 in SNP site；Preferably, described genetic marker combination is included in following 482 SNP sites at least
366；Include at least 405 in following 482 SNP sites as further preferred, described genetic marker combination.It is described
482 SNP sites in US National Biotechnology Information center (National Center for Biotechnology
Information, NCBI) snp database in No. rs be respectively：rs1490413、rs4654468、rs7520386、
rs17351137、rs10927438、rs7517833、rs798066、rs1524180、rs1281611、rs10874326、
rs10801788、rs7546709、rs770927、rs187823、rs6684718、rs2036173、rs10748478、
rs4847034、rs560681、rs10494685、rs7542498、rs922715、rs10495407、rs891700、
rs1413212、rs876724、rs1109037、rs2372009、rs954789、rs863022、rs10203786、
rs12617928、rs13395411、rs6547037、rs12622186、rs4849589、rs11888583、rs1881881、
rs6541835、rs294643、rs993934、rs1251098、rs6711922、rs13383710、rs1540482、
rs10189619、rs12464448、rs2368108、rs12997453、rs1598075、rs13408249、rs850889、
rs1440059、rs2392819、rs1443664、rs4074882、rs907100、rs1357617、rs2030874、
rs4283565、rs4684919、rs1121305、rs4857969、rs4493387、rs4364205、rs9843337、
rs9814482、rs9873254、rs9866013、rs17792804、rs6549661、rs9878912、rs6810199、
rs6437890、rs11921425、rs1436345、rs12639347、rs12488989、rs2399332、rs1872575、
rs6785912、rs1600137、rs150205、rs10936362、rs1384823、rs1017953、rs1355366、
rs779306、rs6444724、rs2046361、rs6846909、rs1356468、rs1452556、rs2324149、
rs11940554、rs6826047、rs11096766、rs1822676、rs279844、rs2570070、rs13151209、
rs13134862、rs6823094、rs2648037、rs10019345、rs11098234、rs1483597、rs167503、
rs619042、rs1948210、rs4473636、rs6813291、rs1554472、rs10024103、rs6838690、
rs35034620、rs6811238、rs921691、rs36029731、rs10520447、rs9684753、rs6552515、
rs1979255、rs7718226、rs11133954、rs717302、rs7714769、rs417513、rs2928231、
rs2257128、rs159606、rs182842、rs10035361、rs10052385、rs641437、rs10072283、
rs767715、rs2406113、rs171227、rs12374491、rs412683、rs12153737、rs10478183、
rs7717565、rs2408032、rs13182883、rs7704770、rs4543310、rs828720、rs315791、
rs154367、rs251934、rs338882、rs1029047、rs1409096、rs13218440、rs9358112、
rs7382170、rs2025679、rs10947859、rs4711791、rs663933、rs9349534、rs2811231、
rs9345749、rs7772342、rs818269、rs12206819、rs16883568、rs6902855、rs9451593、
rs1030428、rs1336071、rs9363138、rs9494677、rs200810、rs12198721、rs7769811、
rs12662867、rs4946582、rs1884255、rs1478829、rs1358856、rs2503107、rs1360947、
rs1452989、rs2272998、rs214955、rs6911233、rs4585574、rs1584535、rs2820408、
rs9347854、rs727811、rs6955448、rs917118、rs1406627、rs10228127、rs1019029、
rs991471、rs6965666、rs7797272、rs7805962、rs9969372、rs9642306、rs10263034、
rs452247、rs6973042、rs1230531、rs10265025、rs10487320、rs1548395、rs321198、
rs737681、rs1861751、rs10092491、rs4237030、rs1377247、rs2368508、rs7831841、
rs1515685、rs1589196、rs7015305、rs7819544、rs7015145、rs4487715、rs10095524、
rs1383474、rs4288409、rs41376144、rs6577791、rs6577808、rs2056277、rs7820230、
rs11781232、rs10092983、rs4606077、rs3858029、rs1015250、rs7042478、rs7049164、
rs10756505、rs1410035、rs2270529、rs10738497、rs1970064、rs7871735、rs1318664、
rs2383542、rs7041158、rs10813060、rs4879342、rs400426、rs7855727、rs11137900、
rs1887719、rs4486277、rs10867685、rs6559615、rs10746789、rs1160209、rs2786797、
rs1327804、rs10818193、rs2781116、rs1335220、rs1463729、rs1360288、rs10776839、
rs962759、rs826472、rs7909413、rs735155、rs7067974、rs7899028、rs3780962、rs7921673、
rs2066058、rs2394137、rs10887346、rs1933957、rs1410059、rs481179、rs1326357、
rs10748937、rs10884478、rs1336414、rs740598、rs1541179、rs748309、rs720416、
rs964681、rs10765075、rs7898450、rs10768550、rs10500617、rs1498553、rs901398、
rs11023546、rs1386734、rs11027682、rs2861329、rs7934283、rs2461678、rs794533、
rs7929050、rs6591147、rs10488710、rs1557451、rs4938230、rs590162、rs4054951、
rs2032414、rs12576089、rs2076848、rs2107612、rs2942582、rs7957282、rs1038666、
rs1909149、rs7980737、rs7299072、rs2171841、rs11836065、rs2111980、rs12812747、
rs10773760、rs1335873、rs1886510、rs9506765、rs528756、rs1538001、rs9315314、
rs7333999、rs2677595、rs9316628、rs6561816、rs2780326、rs1340301、rs359371、
rs7321823、rs9564415、rs9541364、rs4883864、rs9542407、rs337964、rs6562645、
rs495593、rs1146888、rs7323290、rs2783082、rs4885690、rs1215468、rs2794251、
rs1932807、rs7990715、rs9546538、rs9546953、rs2341300、rs12856670、rs9301207、
rs7992038、rs1058083、rs7981712、rs1330527、rs1865350、rs701565、rs6491913、
rs354439、rs9514807、rs1228539、rs1454361、rs1955877、rs1951055、rs2781333、
rs7159343、rs722290、rs2358481、rs4899801、rs2998319、rs10150656、rs9323780、
rs879803、rs873196、rs4530059、rs2016276、rs1821380、rs8037429、rs1528460、
rs9944198、rs4887279、rs7181498、rs16975446、rs2159620、rs729172、rs2342747、
rs7205345、rs4787112、rs12597901、rs16862、rs8056924、rs430046、rs1382387、
rs9905977、rs740910、rs4796362、rs1860555、rs2175957、rs8070085、rs1004357、
rs1027895、rs1897859、rs4080890、rs11077529、rs8081246、rs4793377、rs8078417、
rs2291395、rs4789798、rs689512、rs3744163、rs2292972、rs1493232、rs3906004、
rs16943205、rs1624032、rs9951171、rs7229946、rs273696、rs10502503、rs7232841、
rs12967345、rs2949539、rs12457638、rs1943539、rs985492、rs7240706、rs1567612、
rs1196588、rs9959097、rs7244456、rs4506998、rs17696192、rs521861、rs7243527、
rs1626480、rs1736442、rs154889、rs8092618、rs1906774、rs17176581、rs1244834、
rs1024116、rs719366、rs576261、rs1031825、rs445251、rs12480506、rs2567608、
rs1005533、rs1523537、rs722098、rs2824224、rs2825957、rs2826366、rs2829225、
rs2829292、rs464663、rs2830795、rs2831700、rs2833736、rs914165、rs221956、rs9606186、
rs5746846、rs2073383、rs733164、rs987640、rs5749850、rs2040411、rs1028528、
rs5769866。
The hereditary information of 482 described SNP sites is as shown in table 1：
Table 1
It should be pointed out that above SNP site be the naming method of the snp database according to NCBI representing, the party
Formula be with rs or ac addends position Arabic numerals come represent one determination SNP site.One skilled in the art will appreciate that one
SNP site can also have other representations, such as mark position of certain site on reference to gDNA to represent with HGVS nomenclatures.
The model that the SNP site or SNP site combination referred to as the present invention falls within the present invention is marked using other names mode
Enclose.
The acquisition pattern of above-mentioned 482 SNP sites is：By to NCBI dbGaP data base (http://
Www.ncbi.nlm.nih.gov/gap analysis), have collected wherein 11,989 parts of whole-genome association (Genome-
Wide Association Study, GWAS) the SNP site genotyping result studied, which is balanced through Hardy-Weinberg
After the screening of inspection, 1,842,870 SNP sites are obtained as candidate SNP locus.It is utilized respectively thousand human genome plan (the
1000 Genomes Project) in 853 from different groups independent individuals samples and ten thousand people of Britain plan (the
UK10K project) in 4,000 individual specimen above-mentioned 1, the polymorphism data of 842,870 SNP sites, with equipotential
Gene frequency (Minor Allele Frequency, MAF) is more than or equal to 20% and fixation index (Fixation Index, FRT)
Less than or equal to 0.03 as screening criteria, above-mentioned 1,842,870 SNP site is screened, obtained 12,942 SNP positions
Point；With the r of linkage disequilibrium (LD)2It is screening criteria more than or equal to 0.01, finally obtains 482 SNP positions of the present invention
Point.
Genetic typing is also referred to as genotyping, is to determine idiotype (Genotype) using biological detection method
Technology.Genetic typing is widely used in population genetics and forensic science, be carry out as identity authentication, paternity test,
The powerful of the work such as individual identification, identification of corpse, crime identification, Missing Persons' identification and case investigation.
With the development of round pcr and high throughput sequencing technologies, the method for carrying out genetic typing also develops and perfect, at present
The method of genetic typing mainly has：Direct sequencing, high-flux sequence method, fragment length polymorphism method, fluorescently-labeled fragment
Length polymorphism method, flight mass spectrum detection method, TaqMan fluorescence probe methods, multiple SNaPShot detection methods, ligase detection
Reaction method, improved ligase detection method, gene chips, SNPscan typings, high-resolution melting curve method and it is based on
The SNP typings of qPCR methods.
The sample of genetic typing can be got from any biologic material, for example blood, seminal stain, saliva, tooth, skeleton,
Fingernail, hair, skin, buccal swab, exfoliative cyte and heart, liver, spleen, lung, stomach, kidney, pancreas, brain, intestinal and gallbladder
In organ.
Present invention also offers application of the above-mentioned one group of genetic marker combination in identity authentication.Described application refers to：Profit
The genotype of the SNP site in described genetic marker combination is determined with genetic typing, so as to carry out identity authentication to individuality.
In above-mentioned application, described individuality can be Aisan, American or European, but be not limited to above-mentioned crowd.
In above-mentioned application, the method for described genetic typing is included but is not limited to：Direct sequencing, high-flux sequence method,
Fragment length polymorphism method, fluorescently-labeled fragment length polymorphism method, flight mass spectrum detection method, TaqMan fluorescence probe methods,
Multiple SNaPShot detection methods, Ligase detection reaction method, improved ligase detection method, gene chips, SNPscan point
Type method, high-resolution melting curve method and the SNP typings based on qPCR methods；Preferably high-flux sequence method and gene chip
Method.
In a preferred embodiment of the invention, using high-flux sequence method to containing 482 SNP of the present invention
The genetic marker combination in site carries out genetic typing, can obtain the genotype information of above-mentioned 482 SNP sites simultaneously.
On the other hand, the invention provides a kind of parting kit, described parting kit is comprising for this
SNP site in bright one group of described genetic marker combination carries out the reagent of genetic typing.
Described parting kit is for carrying out identity authentication to individual.
The PCR for the SNP site in one group of genetic marker combination of amplification can also be included in described parting kit
Primer.
Present invention also offers described parting kit is recognized in individual identification, sibship judgement, case investigation, corpse source
Application during fixed, paternity test, criminal are assert and Missing Persons assert.
Present invention also offers a set of compound classification system for identity authentication, described compound classification system is comprising this
The described parting kit of invention and using high-flux sequence method detect instrument needed for SNP site genotype, software, and/or
Module.
In terms of another, present invention also offers a kind of idiotype identity card, described idiotype identity card
Comprising chip, the genotype information of the individual genetic marker site, described genetic marker position on described chip, are stored
Point refers to the SNP site in genetic marker combination of the present invention.
The structural style of described idiotype identity card can be card form, hard-disc type or optical-disk type, but be not limited to
Above-mentioned 3 kinds of structural styles.
Present invention also offers application of the described idiotype identity card in identity authentication.
Compared with prior art, beneficial effects of the present invention are：
(1) one group of genetic marker combination and be combined in classification system that, the present invention is provided, due to the SNP site which includes
Quantity is more, therefore the typing efficiency that can not be compared with the combination of existing genetic marker；Cover comprising each big main population in the whole world,
It is more extensive compared with prior art to the scope of application of different crowd；By entering to the genotype data more than 4000 independent individuals
482 SNP sites that row analysis is obtained, it is relatively reliable compared with prior art to the result of individual identity identification.
(2) a kind of some parents of the one group of genetic marker combination and parting kit that, present invention is provided for blood lineage farther out
Edge relation carries out the effect of family identity authentication and is substantially better than prior art, for some such as grandparents and grandchild of sibship apoplexy due to endogenous wind,
Uncle and nephew, half sib relation carry out the discrimination and accuracy of identity authentication and are substantially better than prior art.
(3) a kind of parting kit and compound classification system that, present invention is provided can be realized to above-mentioned 482 SNP positions
Expand while point, the amplified production comprising 482 SNP sites, convenience and high-efficiency can be obtained by a tube reaction.
(4), typing of the present invention using high throughput sequencing technologies realization to above-mentioned 482 SNP sites, can be obtained simultaneously
The genotype of 482 SNP sites, obtains single individual sequencing library so as to individuality by can realizing a tube reaction
The more succinct high efficient and reliable of identity authentication.
(5) genotype information of individuality, the i.e. present invention, are stored in a kind of idiotype identity card that the present invention is provided
The genotype information of 482 described SNP sites, instrument that can effectively easily as individual identity certification.
(6), one group of genetic marker that the present invention is provided combines and is combined classification system when for identity authentication, true sample
The discrimination of this nothing to do with check sample is excellent, hence it is evident that combine better than str locus seat commonly used in the prior art or SNP site group
Close.It follows that a kind of parting kit of the one group of genetic marker combination provided based on the present invention and a set of compound typing system
Unite when for identity authentication, equally also with the discrimination higher than prior art；Thus, for paternity test, individuality
Identification, identification of corpse, crime are assert, Missing Persons assert and the aspect such as case investigation is with higher than prior art accurate
Property.
Description of the drawings
The likelihood ratio LR comparative results that Fig. 1 is drawn for the combination 2 in experimental example 1.
The likelihood ratio LR comparative results that Fig. 2 is drawn for the combination 2 in experimental example 1.
Specific embodiment
The explanation of following examples is only intended to help and understands the method for the present invention and its core concept.It should be pointed out that right
For those skilled in the art, under the premise without departing from the principles of the invention, the present invention can also be carried out
Some improvement and modification, these improve and modification is also fallen in the protection domain of the claims in the present invention.To disclosed enforcement
The description below of example, enables professional and technical personnel in the field to realize or using the present invention.Various modifications to these embodiments
Will be apparent for those skilled in the art, generic principles defined herein can be without departing from this
In the case of the spirit or scope of invention, realize in other embodiments.Therefore, the present invention is not intended to be limited to illustrated herein
These embodiments in, but can apply to meet the broader model consistent with principles disclosed herein and features of novelty
Enclose.Although can use and heretofore described similar or of equal value any method and material in the enforcement or test of the present invention
Material, place enumerates preferred method and material herein.
Unless otherwise defined, all technologies used herein and scientific terminology with the technical field of the invention
The same meaning that those of ordinary skill is generally understood that.
Unreceipted particular technique or condition person in embodiment, according to technology or condition described by document in the art
(for example write with reference to J. Pehanorm Brookers etc., what Huang Peitang etc. was translated《Molecular Cloning:A Laboratory guide》, the third edition, Science Press) or
Person is carried out according to product description.
The quantitative of embodiment of the present invention amplifying nucleic acid adopts NanoDrop2000 ultramicrospectrophotometers, production code member
ND2000c, purchased from match Mo Feishier companies (Thermo Fisher Scientific)；
GenEluteTMPoba gene group DNA test kit article No. is NA2010-1KT, purchased from SIGMA-ALDRICH companies；
1 one groups of genetic marker combinations of embodiment
Genetic marker combination includes 482 SNP sites of the present invention, No. rs of described 482 SNP sites and
Hereditary information is as shown in table 1.
A kind of 2 parting kit of embodiment
The parting kit is included：
(1), sequencing library reagent preparation；(2), high-flux sequence reagent.
Embodiment 1
Kinsfolk to there are 16 kinships carries out random sample test and interpretation of result.
Individual specimen：Including 16 relatives including child father and mother, brother, sister, cousin, grandparent and grandchild and uncle and nephew etc.
47 kinsfolks altogether of relation；Sample is blood sample.
Experimentation：
To more than, 47 individualities carry out venous blood collection, obtain 47 parts of blood samples, carry out random number to which, carry out double blinding
Experiment：
(1), DNA extraction：
Using GenEluteTMPoba gene group DNA test kit extracts genomic DNA from 47 parts of blood samples respectively, obtains
47 parts of poba gene group DNA samples, concrete extraction process is with reference to test kit operation instruction；
(2), prepared by library：
A, PCR are expanded：
The 47 parts of poba gene group DNA samples for being obtained using the present embodiment step (1) respectively design primer as pcr template,
Using Ion AmpliSeqTM2.0 test kits of Library Kit (are purchased from Life Technologies companies, article No. is Cat.no
4475345) 482 SNP sites in amplified sample, obtain 47 amplified productions；
PCR amplification system：
| Composition | Addition |
| 5×Ion AmpliSeqTMHiFi Mix(red cap) | 4μL |
| Poba gene group DNA | 1ng |
| Amplimer | 10μL |
| Nuclease-free Water | Polishing is to 20 μ L |
PCR amplification programs：
B, using Ion XpressTM1 96Kit test kits of Barcode Adapters are (public purchased from Thermo Fisher
Department, article No. is Cat.no.4474517) respectively 47 amplified productions connection specificitys of the present embodiment step (2)-A acquisitions are connect
Head, obtains 47 different joint-amplified productions, and concrete connection procedure is with reference to test kit operation instruction；
C, useXP Kit test kits (are purchased from Beckman Coulter companies, article No.
For A63880 or A63881) purification is carried out to 47 different joint-amplified productions that the present embodiment step (2)-B is obtained,
After being carried out to joint-amplified production after purification quantitatively using real time fluorescent quantitative nucleic acid amplification detection method, mixing 47 connects
Head-amplified production, constitutes sequencing library；
(3), the sequencing library that the present embodiment step (2)-C is obtained is sequenced using ION PGM sequenators, it is concrete to grasp
Make flow process referring to ION PGM sequenator operating instructions；
(4), interpretation of result：According to the genotyping result of 47 parts of poba gene group DNA samples, sample clustering is carried out at random and is obtained
Individual family relational result, meet the expection sampled before.
At present existing parting kit or for typing genetic marker combination in the present embodiment with regard to grandparent and grandchild and uncle and nephew
The individual family relational result that the individual specimen of relation is obtained, does not meet sampling and is expected.19 traditional str locus are used for example
Seat is carried out to the present embodiment correlated sampless in the individual family relational result that sample clustering is obtained as one group of genetic marker combination,
The sample results of grandparent and grandchild and uncle and nephew relation do not meet the expection sampled before.Illustrate the present invention for for some such as relationships
Grandparent and grandchild, uncle and nephew, half sib relation in relation etc. carry out the effect of identity authentication and are better than prior art.
1 discrimination of experimental example is contrasted
Be utilized respectively traditional 19 str locus seat (hereinafter referred to as combining 1) is combined and using this as genetic marker
482 SNP sites of bright offer carry out identity authentication as genetic marker combination (hereinafter referred to as combining 2) to individuality, and to mirror
Determine result to be compared.
Genetic typing is carried out to combination 1 and combination 2 respectively using high throughput sequencing technologies, and utilizes Elston-Stewart
Algorithm model calculates the likelihood ratio LR of true sibship, and the likelihood ratio LR distributions of its nothing to do with colony are compared.If true
The LR values of real sibship can be noticeably greater than the likelihood ratio LR distributions of unrelated colony, then can carry out the identification of correlation.
As shown in figure 1, transverse axis is likelihood ratio LR scales in figure, two parallel for the 1 likelihood ratio LR comparative results that draw of combination
Vertical line represents the likelihood ratio LR of true sibship, and gray area is the distribution of unrelated colony.Its result cannot separate, and illustrate 19
Identification effect of the individual STR under the sibship is not enough.
The 2 likelihood ratio LR comparative results that draw of combination as shown in fig. 2, it can be seen that the true sibship of its result seemingly
Right rate LR significantly (p value<0.01) more than unrelated population distribution such that it is able to make related identification.
By this experimental example this it appears that the genetic marker that constitutes of 482 SNP sites that the present invention is provided combine for
The effect of identity authentication is substantially better than prior art.
The explanation of above example is only intended to help and understands the method for the present invention and its core concept.It should be pointed out that right
For those skilled in the art, under the premise without departing from the principles of the invention, the present invention can also be carried out
Some improvement and modification, these improve and modification is also fallen in the protection domain of the claims in the present invention.To disclosed enforcement
The described above of example, enables professional and technical personnel in the field to realize or using the present invention.Various modifications to these embodiments
Will be apparent for those skilled in the art, generic principles defined herein can be without departing from this
In the case of the spirit or scope of invention, realize in other embodiments.Therefore, the present invention is not intended to be limited to illustrated herein
These embodiments in, but can apply to meet the broader model consistent with principles disclosed herein and features of novelty
Enclose.
Claims (10)
1. a kind of genetic marker combination, it is characterised in that：During described genetic marker combination includes following 482 SNP sites
At least 214, the rs difference of described 482 SNP sites in the snp database of US National Biotechnology Information center
For：
rs1490413、rs4654468、rs7520386、rs17351137、rs10927438、rs7517833、rs798066、
rs1524180、rs1281611、rs10874326、rs10801788、rs7546709、rs770927、rs187823、
rs6684718、rs2036173、rs10748478、rs4847034、rs560681、rs10494685、rs7542498、
rs922715、rs10495407、rs891700、rs1413212、rs876724、rs1109037、rs2372009、rs954789、
rs863022、rs10203786、rs12617928、rs13395411、rs6547037、rs12622186、rs4849589、
rs11888583、rs1881881、rs6541835、rs294643、rs993934、rs1251098、rs6711922、
rs13383710、rs1540482、rs10189619、rs12464448、rs2368108、rs12997453、rs1598075、
rs13408249、rs850889、rs1440059、rs2392819、rs1443664、rs4074882、rs907100、
rs1357617、rs2030874、rs4283565、rs4684919、rs1121305、rs4857969、rs4493387、
rs4364205、rs9843337、rs9814482、rs9873254、rs9866013、rs17792804、rs6549661、
rs9878912、rs6810199、rs6437890、rs11921425、rs1436345、rs12639347、rs12488989、
rs2399332、rs1872575、rs6785912、rs1600137、rs150205、rs10936362、rs1384823、
rs1017953、rs1355366、rs779306、rs6444724、rs2046361、rs6846909、rs1356468、
rs1452556、rs2324149、rs11940554、rs6826047、rs11096766、rs1822676、rs279844、
rs2570070、rs13151209、rs13134862、rs6823094、rs2648037、rs10019345、rs11098234、
rs1483597、rs167503、rs619042、rs1948210、rs4473636、rs6813291、rs1554472、
rs10024103、rs6838690、rs35034620、rs6811238、rs921691、rs36029731、rs10520447、
rs9684753、rs6552515、rs1979255、rs7718226、rs11133954、rs717302、rs7714769、
rs417513、rs2928231、rs2257128、rs159606、rs182842、rs10035361、rs10052385、
rs641437、rs10072283、rs767715、rs2406113、rs171227、rs12374491、rs412683、
rs12153737、rs10478183、rs7717565、rs2408032、rs13182883、rs7704770、rs4543310、
rs828720、rs315791、rs154367、rs251934、rs338882、rs1029047、rs1409096、rs13218440、
rs9358112、rs7382170、rs2025679、rs10947859、rs4711791、rs663933、rs9349534、
rs2811231、rs9345749、rs7772342、rs818269、rs12206819、rs16883568、rs6902855、
rs9451593、rs1030428、rs1336071、rs9363138、rs9494677、rs200810、rs12198721、
rs7769811、rs12662867、rs4946582、rs1884255、rs1478829、rs1358856、rs2503107、
rs1360947、rs1452989、rs2272998、rs214955、rs6911233、rs4585574、rs1584535、
rs2820408、rs9347854、rs727811、rs6955448、rs917118、rs1406627、rs10228127、
rs1019029、rs991471、rs6965666、rs7797272、rs7805962、rs9969372、rs9642306、
rs10263034、rs452247、rs6973042、rs1230531、rs10265025、rs10487320、rs1548395、
rs321198、rs737681、rs1861751、rs10092491、rs4237030、rs1377247、rs2368508、
rs7831841、rs1515685、rs1589196、rs7015305、rs7819544、rs7015145、rs4487715、
rs10095524、rs1383474、rs4288409、rs41376144、rs6577791、rs6577808、rs2056277、
rs7820230、rs11781232、rs10092983、rs4606077、rs3858029、rs1015250、rs7042478、
rs7049164、rs10756505、rs1410035、rs2270529、rs10738497、rs1970064、rs7871735、
rs1318664、rs2383542、rs7041158、rs10813060、rs4879342、rs400426、rs7855727、
rs11137900、rs1887719、rs4486277、rs10867685、rs6559615、rs10746789、rs1160209、
rs2786797、rs1327804、rs10818193、rs2781116、rs1335220、rs1463729、rs1360288、
rs10776839、rs962759、rs826472、rs7909413、rs735155、rs7067974、rs7899028、
rs3780962、rs7921673、rs2066058、rs2394137、rs10887346、rs1933957、rs1410059、
rs481179、rs1326357、rs10748937、rs10884478、rs1336414、rs740598、rs1541179、
rs748309、rs720416、rs964681、rs10765075、rs7898450、rs10768550、rs10500617、
rs1498553、rs901398、rs11023546、rs1386734、rs11027682、rs2861329、rs7934283、
rs2461678、rs794533、rs7929050、rs6591147、rs10488710、rs1557451、rs4938230、
rs590162、rs4054951、rs2032414、rs12576089、rs2076848、rs2107612、rs2942582、
rs7957282、rs1038666、rs1909149、rs7980737、rs7299072、rs2171841、rs11836065、
rs2111980、rs12812747、rs10773760、rs1335873、rs1886510、rs9506765、rs528756、
rs1538001、rs9315314、rs7333999、rs2677595、rs9316628、rs6561816、rs2780326、
rs1340301、rs359371、rs7321823、rs9564415、rs9541364、rs4883864、rs9542407、
rs337964、rs6562645、rs495593、rs1146888、rs7323290、rs2783082、rs4885690、
rs1215468、rs2794251、rs1932807、rs7990715、rs9546538、rs9546953、rs2341300、
rs12856670、rs9301207、rs7992038、rs1058083、rs7981712、rs1330527、rs1865350、
rs701565、rs6491913、rs354439、rs9514807、rs1228539、rs1454361、rs1955877、
rs1951055、rs2781333、rs7159343、rs722290、rs2358481、rs4899801、rs2998319、
rs10150656、rs9323780、rs879803、rs873196、rs4530059、rs2016276、rs1821380、
rs8037429、rs1528460、rs9944198、rs4887279、rs7181498、rs16975446、rs2159620、
rs729172、rs2342747、rs7205345、rs4787112、rs12597901、rs16862、rs8056924、rs430046、
rs1382387、rs9905977、rs740910、rs4796362、rs1860555、rs2175957、rs8070085、
rs1004357、rs1027895、rs1897859、rs4080890、rs11077529、rs8081246、rs4793377、
rs8078417、rs2291395、rs4789798、rs689512、rs3744163、rs2292972、rs1493232、
rs3906004、rs16943205、rs1624032、rs9951171、rs7229946、rs273696、rs10502503、
rs7232841、rs12967345、rs2949539、rs12457638、rs1943539、rs985492、rs7240706、
rs1567612、rs1196588、rs9959097、rs7244456、rs4506998、rs17696192、rs521861、
rs7243527、rs1626480、rs1736442、rs154889、rs8092618、rs1906774、rs17176581、
rs1244834、rs1024116、rs719366、rs576261、rs1031825、rs445251、rs12480506、
rs2567608、rs1005533、rs1523537、rs722098、rs2824224、rs2825957、rs2826366、
rs2829225、rs2829292、rs464663、rs2830795、rs2831700、rs2833736、rs914165、rs221956、
rs9606186、rs5746846、rs2073383、rs733164、rs987640、rs5749850、rs2040411、
rs1028528、rs5769866。
2. genetic marker combination as claimed in claim 1, it is characterised in that described genetic marker combination includes claim
482 SNP sites described in 1.
3. a kind of parting kit, it is characterised in that：Described parting kit is comprising for claim 1-4 any one
SNP site in described genetic marker combination carries out the reagent of genetic typing.
4. parting kit as claimed in claim 3, it is characterised in that：Described parting kit is also included for expanding power
Profit requires the PCR primer of the SNP site in the genetic marker combination described in 1 or 2.
5. a set of compound classification system, it is characterised in that：Described compound classification system includes dividing described in claim 3 or 4
Type test kit, and using the instrument needed for high-flux sequence method detection SNP site genotype, software, and/or module.
6. a kind of idiotype identity card, it is characterised in that：Described idiotype identity card includes chip, described core
The genotype information of the individual genetic marker site is stored on piece, described genetic marker site is 1 or 2 institute of claim
SNP site in the genetic marker combination stated.
7. the genetic marker described in claim 1 or 2 combines the application in identity authentication.
8. application of the parting kit described in claim 3 or 4 in identity authentication.
9. application of the compound classification system described in claim 5 in identity authentication.
10. application of the idiotype identity card described in claim 6 in identity authentication.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611101246.3A CN106520982B (en) | 2016-12-05 | 2016-12-05 | A kind of compound classification system for identity authentication |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611101246.3A CN106520982B (en) | 2016-12-05 | 2016-12-05 | A kind of compound classification system for identity authentication |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106520982A true CN106520982A (en) | 2017-03-22 |
| CN106520982B CN106520982B (en) | 2019-11-08 |
Family
ID=58355029
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611101246.3A Active CN106520982B (en) | 2016-12-05 | 2016-12-05 | A kind of compound classification system for identity authentication |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106520982B (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106906300A (en) * | 2017-04-21 | 2017-06-30 | 为朔医学数据科技（北京）有限公司 | A kind of genetic ID card and preparation method thereof |
| CN107012226A (en) * | 2017-04-20 | 2017-08-04 | 司法部司法鉴定科学技术研究所 | A kind of detection kit and its detection method of the SNP site based on high-flux sequence |
| CN107868830A (en) * | 2017-11-11 | 2018-04-03 | 深圳深知生物科技有限公司 | A set of SNP site for canine ore grade indexes |
| CN108504744A (en) * | 2018-03-14 | 2018-09-07 | 中国科学院北京基因组研究所 | A kind of micro- haplotype genetic marker and its kit for legal medical expert's detection |
| 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 |
| CN110863056A (en) * | 2018-08-27 | 2020-03-06 | 深圳华大法医科技有限公司 | Method, reagent and application for accurately typing human DNA |
| CN115273976A (en) * | 2022-08-24 | 2022-11-01 | 温州医科大学 | Method, system, equipment and storage medium for identifying half-sib relationship |
| CN115346594A (en) * | 2022-08-24 | 2022-11-15 | 温州医科大学 | Grandfather-grandfather relationship identification method, system, equipment and medium without participation of mother and mother |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102337345A (en) * | 2011-11-04 | 2012-02-01 | 四川大学 | Medicolegal composite assay kit based on twenty triallelic SNP (single nucleotide polymorphism) genetic markers |
| WO2015138997A1 (en) * | 2014-03-14 | 2015-09-17 | Caredx, Inc. | Methods of monitoring immunosuppressive therapies in a transplant recipient |
| CN105483123A (en) * | 2015-11-23 | 2016-04-13 | 元码基因科技（北京）有限公司 | Genetic marker combination, individual gene identity certificate and uses thereof |
-
2016
- 2016-12-05 CN CN201611101246.3A patent/CN106520982B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102337345A (en) * | 2011-11-04 | 2012-02-01 | 四川大学 | Medicolegal composite assay kit based on twenty triallelic SNP (single nucleotide polymorphism) genetic markers |
| WO2015138997A1 (en) * | 2014-03-14 | 2015-09-17 | Caredx, Inc. | Methods of monitoring immunosuppressive therapies in a transplant recipient |
| CN105483123A (en) * | 2015-11-23 | 2016-04-13 | 元码基因科技（北京）有限公司 | Genetic marker combination, individual gene identity certificate and uses thereof |
Non-Patent Citations (1)
| Title |
|---|
| ILLUMINA: "ForenSeq™DNA 特征制备参考指南", 《ILLUMINA》 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107012226A (en) * | 2017-04-20 | 2017-08-04 | 司法部司法鉴定科学技术研究所 | A kind of detection kit and its detection method of the SNP site based on high-flux sequence |
| CN106906300A (en) * | 2017-04-21 | 2017-06-30 | 为朔医学数据科技（北京）有限公司 | A kind of genetic ID card and preparation method thereof |
| CN107868830A (en) * | 2017-11-11 | 2018-04-03 | 深圳深知生物科技有限公司 | A set of SNP site for canine ore grade indexes |
| CN108504744A (en) * | 2018-03-14 | 2018-09-07 | 中国科学院北京基因组研究所 | A kind of micro- haplotype genetic marker and its kit for legal medical expert's detection |
| 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 |
| CN110863056A (en) * | 2018-08-27 | 2020-03-06 | 深圳华大法医科技有限公司 | Method, reagent and application for accurately typing human DNA |
| CN115273976A (en) * | 2022-08-24 | 2022-11-01 | 温州医科大学 | Method, system, equipment and storage medium for identifying half-sib relationship |
| CN115346594A (en) * | 2022-08-24 | 2022-11-15 | 温州医科大学 | Grandfather-grandfather relationship identification method, system, equipment and medium without participation of mother and mother |
| CN115273976B (en) * | 2022-08-24 | 2023-05-05 | 温州医科大学 | Method, system, equipment and storage medium for identifying semi-sibling relation |
| CN115346594B (en) * | 2022-08-24 | 2023-09-05 | 温州医科大学 | Ancestor relationship identification method, system, equipment and medium without raw mother participation |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106520982B (en) | 2019-11-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106520982B (en) | A kind of compound classification system for identity authentication | |
| US11952623B2 (en) | Simultaneous determination of aneuploidy and fetal fraction | |
| US10947595B2 (en) | Nucleic acids and methods for detecting chromosomal abnormalities | |
| US20200385810A1 (en) | Methods for determining fraction of fetal nucleic acids in maternal samples | |
| TWI661049B (en) | Using cell-free dna fragment size to determine copy number variations | |
| Kukurba et al. | RNA sequencing and analysis | |
| EP4043581A1 (en) | Method for generating a paralog assay system | |
| JP2019153332A (en) | Method for determining a copy number variation in sex chromosome | |
| CN105483123B (en) | Genetic marker combination, genes of individuals identity card and application thereof | |
| CN110499373A (en) | Identify the high-throughput STR classification system and kit of complicated affiliation | |
| CN110499372A (en) | Multiplex PCR targeted capture parting system and kit based on high throughput sequencing technologies | |
| Vajpayee et al. | Forensic DNA typing: inception, methodology, and technical advancements | |
| CN105886497A (en) | Allelic ladder of polymorphic short tandem repeat (STR) loci as well as preparation method, identification method and application thereof | |
| WO2018061638A1 (en) | METHOD FOR DETERMINING ORIGIN OF HUMAN GENOMIC DNA OF 100 pg OR LESS THEREFROM, METHOD FOR PERSONAL IDENTIFICATION, AND METHOD FOR ANALYZING DEGREE OF ENGRAFTMENT OF HEMATOPOIETIC STEM CELL | |
| HR MAS et al. | From DNA to Data: Transcending Beyond the Double Helix and Demystifying the Genetic Alchemy of Life Through NGS to Empower Precision Medicine | |
| Jain et al. | Technologies & Applications | |
| SOHTORİK ÖZTÜRK et al. | Determination of Gene Frequency of 10 New STR Loci in Turkey: Experimental Research. | |
| CN117947178A (en) | Genetic marker combination and composite typing system for family identity identification | |
| US20130023427A1 (en) | Methods for assessing genomic instabilities in tumors |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |