CN105335625A - Genetics detecting device of embryo before implantation - Google Patents

Abstract

The invention relates to the field of biology, and provides a genetics detecting device of an embryo before implantation. The genetics detecting device of the embryo before implantation comprises a message unit 100, an amphiphilic haplotype creating module 200, a candidate embryo haplotype building module 300, a candidate embryo haplotype parental source determining module 400 and a judgment module 500 for judging whether a candidate embryo carries gene defects. According to the genetics detecting device of the embryo before implantation provided by the invention, a more accurate genetics diagnosis of the embryo before implantation can be given, so as to select out healthy embryos.

Description

Science of heredity pick-up unit before Embryonic limb bud cell

Technical field

The present invention relates to field of biology, particularly relate to the science of heredity pick-up unit before a kind of Embryonic limb bud cell.

Background technology

Hereditary disease not only has a strong impact on the physical and mental health of patient, bring very large burden to society and family, and treatment is very difficult, even if can treat, expense also costly, therefore preimplantation genetic diagnosis can avoid the birth of hereditary disease fetus, to improving the health of the people, alleviates burden on society significant.

Preimplantation genetic diagnosis technology (PreimplantationGeneticDiagnosis, PGD) be on the basis of supplementary reproduction and molecular biology fast development, mutually merge the new diagnostic techniques of of being formed, the history of existing more than 20 year so far.Preimplantation genetic diagnosis is an extraordinary measure for the treatment of improve the health of the people, control inborn defect and infertility.2010, " father of test-tube baby " British scientist RobertEdwards obtained the affirmative of Nobel prize's soul.Tube-test baby techniques is as a far-reaching technology, facilitate Issues of Human Assisted Reproductive Technologies (assistedreproductivetechnology, ART) producing and developing, has also driven the scientific research of reproductive medicine association area and the development of clinical practice.From 1978, first case test-tube baby was born in the world, so far to be born about 4,000,000 people by this technology in the whole world, some of them have had themselves offspring, and this technology experienced by three crucial developing stage between these three more than ten years, defines successive three generations's auxiliary procreation technology.First generation auxiliary procreation technology, be also referred to as technology (invitrofertilization in vitro fertilization, IVF), refer to by B ultrasonic or endoscope, in woman's ovary, take out ovum, and in test tube or double dish, cultivate fertilization together with sperm, after embryonated egg continuation is trained embryo, again by this embryo transfer in the uterus of parent, embryo nidation in maternal uterine is also continued to grow.This technology usually can solve Female Primary because of sterility disease, as salpingemphraxis; Second generation auxiliary procreation technology, be also referred to as intracytoplasmic sperm injection technology (intra-cytoplasmicsperminjection, or Microfertilization technology ICSI), the feature of ICSI technology utilizes micromanipulation exactly, has artificially carried out sperm ovum binding by single sperm injection to oocyte cytoplasm.It is sterile that second-generation technology mainly solves that male sex's reason causes, as few essence/azoospermatism; Third generation auxiliary procreation technology, be otherwise known as preimplantation genetic diagnosis (preimplantationgeneticdiagnosis, or examination (preimplantationgeneticscreening PGD), PGS), genetic diagnosis technology is mainly utilized to ensure the hereditary well-being of offspring.Preimplantation genetic diagnosis (PreimplantationGeneticDiagnosis, PGD) be at Issues of Human Assisted Reproductive Technologies (AssitedReproductiveTechnology, ART) basis is carried out a diagnostic techniques of genetic analysis to gamete or IVF Embryos, object is the Mr. and Mrs for there being genetic defect, get rid of the gamete or the embryo that there are genetic defect, fetal tissues is selected to transplant, meet the hope that they give birth to normal offspring, reduce gestation and the birth of hereditary disease infant.PGD is applied to clinical indication and comprises: sex-kink disease, single gene inheritance disease, chromosome structure and numerical abnormality and the Mr. and Mrs having the birth of excessive risk hereditary disease infant to be inclined to.For having the accurate father and mother of hereditary disease family history and having given birth to the father and mother that is suffered from hereditary disease child, have serious hereditary disease risk when giving birth to child, now answer is accurately and timely all very important.

At present, the conventional PGD technology for chromosome genetics examination mainly contains fluorescence in situ hybridization technique, comparative genomic hybridization hybrid chip technology and mononucleotide polymorphism chip technology.The limitation of FISH technology is mainly manifested in due to number of probes quantitative limitation, and FISH technology cannot the whole 23 pairs of chromosomes of examination, cannot carry out complete detection to chromosomal rearrangement and chromosome aneuploid.The limitation of aCGH technology is mainly manifested in the normal genome rearrangement of None-identified copy number or chromosome aberration, cannot distinguish fetal tissues and balanced translocation embryo; Can not directly read chromosome deficiency and repetition, need could differentiate that embryo chromosome doubly sexually revises and uniparental disomy in conjunction with direct heredity information.Single nucleotide polymorphism gene (singlenucleotidepolymorphism, SNP) resolution of chip technology is up to 1.5kb, far above other detection techniques, the non-equilibrium chromosome translocation of the micro-segments that above several method is undetected, repetition and disappearance can be found.The advantage of this technology is that the exception that not only can detect aneuploid also can detect euploid extremely simultaneously, and by analyzing special single nucleotide site, also can the phenomenon such as examination monogenic disease and uniparental disomy.Simultaneously also supplementary reproduction other in also have and apply more widely.In the PGS of aneuploid, SNP chip technology obtains certain affirmative.By analyzing the genotype of embryo and the haplotype of parents, the haplotype of embryo can be obtained.Up to the present, on the one hand, unicellular gene magnification still can not avoid the problem of allele dropout (Alleledrop-out, ADO) completely.ADO is because very few one of them the caused allele of test material does not increase, or amplification quantity not sufficient is to reach the level of detecting, he can cause the erroneous judgement of PGD result, directly can cause the implantation of an embryo in extreme circumstances, cause serious consequence, therefore, overcome the erroneous judgement that ADO causes and just become very crucial; On the other hand, the method for existing analysis embryo haplotype is also very complicated, easily makes mistakes.

Summary of the invention

The shortcoming of prior art in view of the above, the object of the present invention is to provide the science of heredity pick-up unit before a kind of Embryonic limb bud cell and application thereof.

Science of heredity pick-up unit before Embryonic limb bud cell of the present invention comprises:

Message unit 100, for obtaining the SNP site information of candidate embryo sample father, the SNP site information of candidate embryo sample mother, candidate embryo sample father and mother's phenotypic information, the SNP site information of propositus, the SNP site information of candidate embryo sample, the individual information of propositus, and store single gene inheritance disease information;

Amphiphilic monomer type creation module 200, be connected with SNP site message unit 100, the individual information for the SNP site information of the SNP site information of the SNP site information according to candidate embryo sample father, candidate embryo sample mother, propositus, single gene inheritance disease information and propositus is established: the target SNP site combination of the target gene that amphiphilic monomer type is corresponding; Two chromosomal equipotential types of each target SNP site parents thus build amphiphilic monomer type.

Candidate embryo haplotype reconstruction module 300, be connected with amphiphilic monomer type creation module 200 and message unit 100, determine for the SNP site information of the SNP site information of the SNP site information according to candidate embryo sample father, candidate embryo sample mother, candidate embryo sample, the target SNP site combination of target gene that amphiphilic monomer type is corresponding, two chromosomal equipotential types of each target SNP site parents: the crucial SNP site combination of the target gene that candidate embryo haplotype is corresponding, each crucial SNP site two chromosomal parental sources in candidate embryo.

Candidate embryo haplotype parental source determination module 400, be connected with candidate embryo haplotype reconstruction module 300, for the crucial SNP site combination according to target gene corresponding to candidate embryo haplotype, and each crucial SNP site two chromosomal parental sources are determined: the parental source of target gene candidate embryo haplotype.

Whether candidate embryo carries gene defect judge module 500, be connected with candidate embryo haplotype parental source determination module 400 and message unit 100, parental source, judgement for two chromosomal haplotypes of the candidate embryo according to single gene inheritance disease information, propositus's individual information, candidate embryo sample father and mother's phenotypic information, target gene: whether candidate embryo carries the gene defect in parents.

As further preferred, the science of heredity pick-up unit before Embryonic limb bud cell of the present invention also comprises:

Candidate's embryo chromosome abnormality juding module 600, is connected with message unit 100, for according to the SNP site information of candidate embryo sample and the SNP site information comparison of normal sample, marks the region having chromosome abnormality in candidate embryo.

Same conduct is further preferred, and message unit 100 also provides HLA related gene information, and the science of heredity pick-up unit before Embryonic limb bud cell also comprises:

Candidate embryo HLA distribution type analysis module 700, be connected with message unit 100 and candidate embryo haplotype parental source determination module 400, for with HLA related gene for target gene, according to the parental source of target gene candidate embryo haplotype, filter out the candidate embryo consistent with the parental source of propositus's target gene.

Same conduct is further preferred, and the science of heredity pick-up unit before Embryonic limb bud cell also comprises:

Sample Quality Control unit 800, be connected with message unit 100, for judging whether the recall rate of the SNP of candidate embryo sample father, the SNP site of candidate embryo sample mother, the SNP site of propositus, the SNP site of candidate embryo exceedes threshold value, and judge whether the sex input of candidate embryo sample parents' sex and propositus is wrong.

For amphiphilic monomer type creation module 200, preferably, can comprise further:

The target SNP site combination determination module 210 of target gene, be connected with message unit 100, for for target gene target setting region, determine according to the SNP site information of candidate embryo sample father in target area, the SNP site information of candidate embryo sample mother, single gene inheritance disease information: the target SNP site combination of the target gene that target gene amphiphilic monomer type is corresponding.

Two chromosomal equipotential determination type module 220 of each target SNP site parents, determination module 210 is combined and SNP site message unit 100 is connected with the target SNP site of target gene, for utilizing the SNP site information of candidate embryo sample father, the SNP site information of candidate embryo sample mother, the SNP site information of propositus, family's genealogical relationship between propositus and candidate embryo, the target SNP site combined information of target gene, determine: in the target SNP site combination of described target gene, the equipotential type of each target SNP site on two chromosomes of candidate embryo parents, thus build amphiphilic monomer type.

For candidate embryo haplotype parental source determination module 400, preferably, comprise further:

Candidate embryo haplotype parental source statistical module 410, be connected with candidate embryo haplotype reconstruction and genetic origin determination module 300 and amphiphilic monomer type creation module 200, for the crucial SNP site combination corresponding according to target gene candidate embryo haplotype, and each crucial SNP site two chromosomal parental sources, in statistics target gene candidate embryo haplotype, the crucial SNP site sum that all kinds of crucial SNP site two chromosomal parental sources are corresponding.

Candidate embryo haplotype parental source analysis module 420, be connected with candidate embryo haplotype parental source statistical module 410, for the crucial SNP site sum according to all kinds of parental source of target gene candidate embryo haplotype, determine candidate embryo haplotype parental source.

The present invention, detecting on the basis of data based on SNP chip, utilizes the science of heredity pick-up unit before Embryonic limb bud cell of the present invention, avoid the result erroneous judgement that allele dropout problem causes, and analytic process is more succinct, not easily makes mistakes.The SNP chip result of science of heredity pick-up unit before Embryonic limb bud cell of the present invention to parents, propositus and embryo carries out automated analysis, quick output result, thus get rid of the embryo having genetic defect, fetal tissues is selected to transplant, meet the hope of the normal offspring of fertility, reduce gestation and the birth of hereditary disease infant.User only needs single job can complete the testing of whether carrying gene defect of a large amount of embryo, disposablely can complete the detection of HLA distribution type simultaneously, thus the procedure operation time of greatly having saved needed for user, also help data analyst to increase work efficiency simultaneously.

Accompanying drawing explanation

Fig. 1 is the schematic diagram of the device of the embodiment of the present invention

Fig. 2 is the schematic diagram of the device of one embodiment of the present invention

Fig. 3 is the schematic diagram of the device of another preferred embodiment of the present invention

Fig. 4 is the schematic diagram of the device of the another preferred embodiment of the present invention

Fig. 5 is the schematic diagram of the device of the another preferred embodiment of the present invention

Fig. 6 is the schematic diagram of the device of most preferred embodiment of the present invention

Embodiment

The present invention is based on gene and genomic level, before Embryonic limb bud cell, carry out science of heredity detection.At present, science of heredity before Embryonic limb bud cell detects and comprises the cause a disease Carriage of haplotype (being the haplotype carrying gene defect) of single-gene disorder and detect and chromosome abnormality detection.The present invention first for single-gene disorder cause a disease haplotype Carriage detect improve, the cause a disease Cleaning Principle of Carriage of haplotype of single-gene disorder of the present invention is: if single-gene disorder causes a disease haplotype in embryo genetic parents, then when SNP chip detects, embryo's haplotype of this target gene regions is the same with haplotype pathogenic in parents.

In the present invention, detecting embryo, whether to carry pathogenic haplotype be exactly contrast by propositus and father and mother's SNP site, in conjunction with family's genealogical relationship of propositus and father and mother, to determine the acquired haplotype of target gene regions parents, then contrast the SNP site of embryo and father and mother, in conjunction with family's genealogical relationship of embryo and propositus, judge the genetic origin of the haplotype of embryo chromosome, again in conjunction with the mode of inheritance of single gene inheritance disease, thus the Carriage of the pathogenic haplotype of clear and definite embryo.

The present invention also can comprise chromosome abnormality further and detect.Abnormal chromosome detects the chromosomal region just referring to and find and change.Detect chromosome abnormality, Cleaning Principle of the present invention is: B gene frequency band exists great difference compared with normal chromosomal, and the analysis by changing B gene frequency band judges that whether the chromosome of embryo is abnormal.

If the chromosome abnormality of embryo, then when SNP chip detects, the band number of the B gene frequency figure in this region will change, concrete:

1) chromosome number increases, and the number being embodied in band in B gene frequency figure increases

2) chromosome number reduces, and is embodied in the decreased number of band in B gene frequency figure.

The present invention, detecting based on SNP chip on the basis of data, by appropriate technical finesse, avoids the result erroneous judgement that allele dropout problem causes, can provide diagnostic result more accurately, to pick out healthy embryo.

Below by way of specific instantiation, embodiments of the present invention are described, those skilled in the art the content disclosed by this instructions can understand other advantages of the present invention and effect easily.The present invention can also be implemented or be applied by embodiments different in addition, and the every details in this instructions also can based on different viewpoints and application, carries out various modification or change not deviating under spirit of the present invention.

In addition should be understood that the one or more method steps mentioned in the present invention do not repel and can also to there is additive method step or can also insert additive method step before and after described combination step between these steps clearly mentioned, except as otherwise noted; Will also be understood that, the relation that is connected between the one or more equipment/devices mentioned in the present invention is not repelled and can also to be there are other equipment/devices or can also insert other equipment/devices before and after described unit equipment/device between these two equipment/devices clearly mentioned, except as otherwise noted.And, except as otherwise noted, the numbering of various method steps is only the convenient tool differentiating various method steps, but not be ordering or the enforceable scope of restriction the present invention of restriction various method steps, the change of its relativeness or adjustment, when changing technology contents without essence, when being also considered as the enforceable category of the present invention.

In the present invention, each letter related in formula and sign only represent for convenience of the symbol of record and explanation, and as required, those skilled in the art also can adopt other symbols to represent.

Fig. 1 is the structural scheme of mechanism of the science of heredity pick-up unit before Embryonic limb bud cell of the present invention.

As shown in the figure, the science of heredity pick-up unit before Embryonic limb bud cell of the present invention comprises:

Message unit 100, for obtaining the SNP site information of candidate embryo sample father, the SNP site information of candidate embryo sample mother, candidate embryo sample father and mother's phenotypic information, the SNP site information of propositus, the SNP site information of candidate embryo sample, the individual information of propositus, and store single gene inheritance disease information;

Amphiphilic monomer type creation module 200, be connected with SNP site message unit 100, the individual information for the SNP site information of the SNP site information of the SNP site information according to candidate embryo sample father, candidate embryo sample mother, propositus, single gene inheritance disease information and propositus is established: the target SNP site combination of the target gene that amphiphilic monomer type is corresponding; Two chromosomal equipotential types of each target SNP site parents thus build amphiphilic monomer type.

Candidate embryo haplotype reconstruction module 300, be connected with amphiphilic monomer type creation module 200 and message unit 100, determine for the SNP site information of the SNP site information of the SNP site information according to candidate embryo sample father, candidate embryo sample mother, candidate embryo sample, the target SNP site combination of target gene that amphiphilic monomer type is corresponding, two chromosomal equipotential types of each target SNP site parents: the crucial SNP site combination of the target gene that candidate embryo haplotype is corresponding, each crucial SNP site two chromosomal parental sources in candidate embryo.

Candidate embryo haplotype parental source determination module 400, be connected with candidate embryo haplotype reconstruction module 300, for the crucial SNP site combination according to target gene corresponding to candidate embryo haplotype, and each crucial SNP site two chromosomal parental sources are determined: the parental source of target gene candidate embryo haplotype.

Whether candidate embryo carries gene defect judge module 500, be connected with candidate embryo haplotype parental source determination module 400 and message unit 100, parental source, judgement for two chromosomal haplotypes of the candidate embryo according to single gene inheritance disease information, propositus's individual information, candidate embryo sample father and mother's phenotypic information, target gene: whether candidate embryo carries the gene defect in parents.

In a preferred embodiment, as shown in Figure 2, the science of heredity pick-up unit before Embryonic limb bud cell also comprises:

Candidate's embryo chromosome abnormality juding module 600, is connected with message unit 100, for according to the SNP site information of candidate embryo sample and the SNP site information comparison of normal sample, marks the region having chromosome abnormality in candidate embryo.

In another preferred embodiment, as shown in Figure 3, message unit 100 also provides HLA related gene information, and the science of heredity pick-up unit before Embryonic limb bud cell also comprises:

Candidate embryo HLA distribution type analysis module 700, be connected with message unit 100 and candidate embryo haplotype parental source determination module 400, for with HLA related gene for target gene, according to the parental source of target gene candidate embryo haplotype, filter out the candidate embryo consistent with the parental source of propositus's target gene.

In another preferred embodiment, as shown in Figure 4, the science of heredity pick-up unit before Embryonic limb bud cell also comprises:

Sample Quality Control unit 800, be connected with message unit 100, for judging whether the recall rate of the SNP of candidate embryo sample father, the SNP site of candidate embryo sample mother, the SNP site of propositus, the SNP site of candidate embryo exceedes threshold value, and judge whether the sex input of candidate embryo sample parents' sex and propositus is wrong.

Launch to describe in detail with regard to unit below:

Message unit 100

Each SNP site information at least comprises: the chromosome position of SNP, base composition and genotype information.

Obtain SNP information from test sample book can adopt chip detection or utilize second generation sequencing technologies to obtain rear import information unit 100, preferred chip detecting method.Described chip detection is undertaken by any chip method, includes but not limited to Illumina optical fiber superbead chip technology.The HumanCytoSNP-12BeadChip that detection chip that SNP information uses derives from Illumina company is obtained in embodiment.

Single gene inheritance disease information at least comprises: the mode of inheritance information of the gene information that single gene inheritance disease is corresponding, single gene inheritance disease.

The gene information that single gene inheritance disease is corresponding can be the related gene information such as the gene location that single gene inheritance disease is corresponding.These information can obtain according to prior art.

In the mode of inheritance information of single gene inheritance disease, the mode of inheritance of described single gene inheritance disease is selected from: autosomal dominant inheritance, autosomal recessive inheritance, X sexlinked dominant inheritance and x linked recessive heredity.These information can also obtain according to prior art.

Propositus: refer to be made a definite diagnosis the people suffering from this single-gene disorder in parents' family.Propositus can be the child of candidate embryo father and mother, can also be siblings and the father and mother thereof of candidate father embryo, also can be siblings and the father and mother thereof of candidate mother embryo.

Propositus of the present invention is divided into children propositus, paternal propositus and maternal propositus.If propositus is the child of candidate embryo father and mother, then this propositus is children propositus; If propositus is siblings and the father and mother thereof of candidate father embryo, then this propositus is paternal propositus; If propositus is siblings and the father and mother thereof of candidate mother embryo, then this propositus is maternal propositus.

The individual information of propositus at least comprises: family's genealogical relationship of propositus and candidate embryo.

Family's genealogical relationship of propositus and candidate embryo can be selected from: elder brother, elder sister, grandfather, grandmother, aunt, uncle, granddad, grandmother, aunt or uncle.

Elder brother described in the present invention, elder sister are the children of candidate embryo father and mother, grandfather, grandmother are the father and mother of candidate father embryo, aunt, uncle are the blood brother sister of candidate father embryo, granddad, grandmother are the father and mother of candidate mother embryo, and aunt, uncle are the blood brother sister of candidate mother embryo.

Propositus can be son or the daughter of these father and mother, and the family's genealogical relationship so between propositus and candidate embryo is exactly elder brother or elder sister; Propositus also can be the father of this father, mother, sister or brother, and the family's genealogical relationship so between propositus and candidate embryo is exactly grandfather, grandmother, aunt or uncle; Propositus also can be the father of this mother, mother, sister or brother, and the family's genealogical relationship so between propositus and candidate embryo is exactly granddad, grandmother, aunt, uncle.

Amphiphilic monomer type creation module 200

Target gene is gene to be analyzed, can be the Disease-causing gene that single gene inheritance disease to be analyzed is corresponding, also can be other interested genes, such as HLA related gene.

In the present invention, haplotype refers to: one group of specific region interrelated, and the SNP site tending to entail with entirety offspring is combined in equipotential type combination corresponding on item chromosome.From upper concept, haplotype all has the SNP site combination of its correspondence.If determine the equipotential type of each SNP site in SNP site combination corresponding to haplotype, this haplotype can be built into.

Equipotential type: in the present invention, base A and base T is defined as A type equipotential, and base C and bases G are defined as Type B equipotential.

For example, one group 8 the SNP site combinations that are mutually related, the base that each SNP site is corresponding on item chromosome is respectively C, T, A, T, G, G, A and C, the equipotential type that then on this chromosome, each SNP site is corresponding is respectively B, A, A, A, B, B, A and B, and so can build and obtain this SNP site haplotype be combined on this chromosome is BAAABBAB.

SNP site: refer to SNP position on chromosome.Because human chromosome occurs in pairs, therefore corresponding two the equipotential types of SNP site.

Acquired haplotype: in the present invention, by on autosome or X chromosome, candidate embryo parental chromosomes has with propositus on the haplotype of genetic association and candidate's embryo chromosome and has the haplotype of genetic association to be all appointed as acquired haplotype with propositus.In the present invention, described acquired haplotype can be the haplotype that single gene inheritance disease gene pairs is answered, and also can be the haplotype that HLA distribution type gene pairs is answered.

Genetic association is had to refer to: to have genetic affinity.

For haplotype.Based on genetic principle, human chromosome exists in pairs, and haplotype also exists in pairs, and one of them is derived from father, and another is derived from mother.If grandfather has a pair haplotype g1 and g2, haplotype corresponding to father is h1 and h2, wherein h1 heredity is from g1, the haplotype of brother's correspondence of father is i1 and i2, wherein i1 heredity is from g1, the haplotype of children one correspondence of father is j1 and j2, wherein j1 heredity is from h1, the haplotype of children two correspondence of father is k1 and k2, wherein k1 heredity is from h2, due to h1, i1, j1 is all directly or indirectly hereditary from g1, so think grandfather, father and brother and children in only have haplotype g1, h1, i1, genetic affinity is had between j1, i.e. haplotype g1, h1, i1, j1 has genetic association, without genetic affinity between all the other haplotypes, namely between all the other haplotypes without genetic association.Can in like manner analogize for SNP site.

In the present invention, target gene amphiphilic monomer type refers to: the target SNP site of target gene is combined in equipotential type combination corresponding respectively on candidate father embryo two chromosomes and candidate mother embryo two chromosomes.From upper concept, target gene amphiphilic monomer type has the target SNP site combination of the target gene of its correspondence.In target gene amphiphilic monomer type, the haplotype be positioned on candidate father embryo two chromosomes is denoted as p1 and p2 respectively, is positioned at the haplotype on candidate mother embryo two chromosomes, is denoted as m1 and m2 respectively.

Genotype: in the present invention, is called genotype two equipotential types of position a certain on chromosome.In the case, the genotype of any one SNP site only has three kinds of situations, and AA type, AB type, BB type, and wherein AA type and BB type are homozygous, and AB type is heterozygous.For the SNP site on male X chromosome, if the equipotential type on X chromosome is A, then its genotype is designated as homozygous AA type; If the equipotential type on X chromosome is B, then its genotype is designated as homozygous BB type.When ADO occurs autosome a certain SNP site, the genotypic markers in this site is the homozygous of the equipotential type do not lost.If autosomal two equipotential types are AB originally, after there is ADO, lost an equipotential B, only remaining A, then its genotypic markers is homozygous AA.

Two chromosomal equipotential types of target SNP site parents comprise: be positioned at the equipotential type on two chromosomes of candidate father embryo, be denoted as P respectively _1gtwith P _2gt; Be positioned at the equipotential type on two chromosomes of candidate mother embryo, be denoted as M respectively _1gtwith M _2gt.

Further, the chromosomal equipotential type P of target SNP site two _1gtand P _2gtin, P _1gtrefer to the equipotential type of target SNP site in haplotype p1, P _2gtrefer to the equipotential type of target SNP site in haplotype p2.

The chromosomal equipotential type M of target SNP site two _1gtand M _2gtin, M _1gtrefer to the equipotential type of target SNP site in haplotype m1, M _2gtrefer to the equipotential type of target SNP site in haplotype m2.

Target SNP site (i.e. tagSNP) in described target SNP site combination can be one or more.Be generally by multiple interrelated, tend to the combination that target SNP site (i.e. tagSNP) that entirety entails offspring is formed.In particular cases, screen less than more target SNP site, also only can set up haplotype according to single target SNP site.Generally, in the combination of target SNP site, target SNP site number is more, and credible result degree is higher.

Target SNP site is divided into the effective tagSNP of father and the effective tagSNP of mother.

The effective tagSNP of father: exist only on autosome, the genotype (P of father in a certain SNP site _gt) for heterozygous (AB) and the genotype (M of mother _gt) be homozygous (AA or BB), so this site is the effective tagSNP of father.The effective tagSNP of father can be used for the equipotential type P determining that this target SNP site is corresponding in father two haplotype p1 and p2 _1gtand P _2gt.

The effective tagSNP of mother: may reside on autosome and also may reside on X chromosome, for autosome, the genotype (M of mother in a certain SNP site _gt) for heterozygous (AB) and the genotype (P of father _gt) be homozygous (AA or BB), so this site is the effective tagSNP of mother; For X chromosome, as long as the genotype (M of mother in a certain SNP site _gt) be heterozygous, so this site is the effective tagSNP of mother.The effective tagSNP of mother can be used for the equipotential type M determining that this target SNP site is corresponding in mother two haplotype m1 and m2 _1gtand M _2gt.

In one particular embodiment of the present invention, as shown in Figure 5, amphiphilic monomer type creation module 200 comprises further:

The target SNP site combination determination module 210 of target gene, be connected with message unit 100, for for target gene target setting region, determine according to the SNP site information of candidate embryo sample father in target area, the SNP site information of candidate embryo sample mother, single gene inheritance disease information: the target SNP site combination of the target gene that target gene amphiphilic monomer type is corresponding.

Two chromosomal equipotential determination type module 220 of each target SNP site parents, determination module 210 is combined and SNP site message unit 100 is connected with the target SNP site of target gene, for utilizing the SNP site information of candidate embryo sample father, the SNP site information of candidate embryo sample mother, the SNP site information of propositus, family's genealogical relationship between propositus and candidate embryo, the target SNP site combined information of target gene, determine: in the target SNP site combination of described target gene, the equipotential type of each target SNP site on two chromosomes of candidate embryo parents, thus build amphiphilic monomer type.

Form in the target SNP site composite module 210 of target gene amphiphilic monomer type,

Described target area is region shared by target gene; Or be the overall area shared by target gene and target gene upstream and downstream certain area.

If target gene length is enough, and tagSNP existing suitable in this gene region, then target area is only region shared by target gene.If the length of target gene is shorter, suitable tagSNPs is not had in target gene regions, if so only with region shared by target gene for target area, just have no idea to create the haplotype of parents, in this case, when analyzing, choosing target gene and upstream and downstream certain area and building amphiphilic monomer type as target area.The not specific regulation of described target gene upstream and downstream certain area, can find at least one suitable tagSNP for benchmark.Within generally caning be controlled in target gene upstream and downstream 5Mb, within preferred 2MB.

According to the definition of preceding aim SNP site, the effective tagSNP of the father in target area and the effective tagSNP of mother can be found, thus obtain the target SNP site combination of target gene.

For two chromosomal equipotential determination type module 220 of each target SNP site parents.

In one embodiment of the invention, the equipotential type that following formula is determined on two chromosomes of candidate embryo parents is adopted:

Alphabetical implication in formula:

P _1gt: the equipotential type referring to target SNP site in haplotype p1 on autosome or X chromosome, its value can be A or B;

P _2gt: the equipotential type referring to target SNP site in haplotype p2 on autosome, its value can be A or B.Be positioned at the target SNP site on X chromosome, then there is not P _2gt;

M _1gt: the equipotential type referring to target SNP site in haplotype m1 on autosome or X chromosome, its value can be A or B;

M _2gt: the equipotential type referring to target SNP site in haplotype m2 on autosome or X chromosome, its value can be A or B;

P1: refer to the target gene haplotype on the item chromosome of candidate embryo sample father; P2: refer to the target gene haplotype on the another item chromosome of candidate embryo sample father; When there is acquired haplotype in p1 and p2, p1 is specified to be acquired haplotype; When target gene is positioned on X chromosome, p2 does not exist.

M1: refer to the target gene haplotype on the item chromosome of candidate embryo sample mother; M2: refer to the target gene monomer on the another item chromosome of candidate's Embryos Embryo sample mother; When there is acquired haplotype in m1 and m2, m1 is specified to be acquired haplotype;

R _gt: the genotype referring to target SNP site propositus, its value is AA, AB or BB, and its value derives from message unit.

P _gt: the genotype referring to target SNP site candidate embryo sample father, its value is AA, AB or BB, and its value derives from message unit.

M _gt: the genotype referring to target SNP site candidate embryo sample mother, its value is AA, AB or BB, and its value derives from message unit.

A: the base-pair referred in target SNP site is the equipotential type of base A and base T;

B: the base-pair referred in target SNP site is the equipotential type of base C and bases G;

Should be understood that each designations is only statement and conveniently sets up, on the basis that implication is constant, other schematic symbols also can be adopted to replace.

1) between propositus and candidate embryo, family's genealogical relationship is elder brother or elder sister, when target gene is on autosome,

As follows to father effective tagSNP site computing formula:

$M_{1gt}=M_{2gt}=\left\{\begin{array}{c}A\left(M_{gt}=AA\right)\\ B\left(M_{gt}=BB\right)\end{array}\right.---\left(1\right)$

P _1gt＝R _gt-M _1gt(a2)

P _2gt＝P _gt-P _1gt(3)

Formula (1) represents, the genotype M of target SNP site mother _gtduring for AA, M _1gtwith M _2gtequal value is A; The genotype M of target SNP site mother _gtduring for BB, M _1gtwith M _2gtequal value is B;

Formula (a2) represents, asks R _gtwith M _1gtdifference set as P _1gtvalue.By R _gt(being made up of two equipotential types) removes M _1gtequipotential type after the equipotential type represented under complementation is appointed as P _1gtvalue.

Formula (3) represents, asks P _gtwith P _1gtdifference set as P _2gtvalue.By P _gt(being made up of two equipotential types) removes P _1gtequipotential type remaining after the equipotential type represented is as P _2gtvalue.In formula (3), P _gtvalue is AB;

Think under this situation, p1 and m1 is acquired haplotype, all non-acquired haplotype of p2 and m2.

For the effective tagSNP site of father on autosome, the genotype of this site father is AB, and the genotype of mother is AA or BB.In one embodiment, the genotype of mother is AA, and propositus's genotype is AB, so M _1gtwith M _2gtbe A, R _gtwith P _gtbe AB, P _1gt=R _gt-M _1gt=AB-A=B; P _2gt=P _gt-P _1gt=AB-B=A; In another embodiment, the genotype of mother is BB, and propositus's genotype is AB, so M _1gtwith M _2gtbe B, R _gtwith P _gtbe AB, P _1gt=R _gt-M _1gt=AB-B=A; P _2gt=P _gt-P _1gt=AB-A=B; In another embodiment, the genotype of mother is BB, and propositus's genotype is BB, M _1gtwith M _2gtbe B, R _gtfor BB, P _gtfor AB, P _1gt=R _gt-M _1gt=BB-B=B; P _2gt=P _gt-P _1gt=AB-B=A.

Mother effective tagSNP site computing formula is as follows:

$P_{\lg t}=P_{2gt}=\left\{\begin{array}{c}A\left(P_{gt}=AA\right)\\ B\left(P_{gt}=BB\right)\end{array}\right.---\left(a4\right)$

M _1gt＝R _gt-P _1gt(a5)

M _2gt＝M _gt-M _1gt(6)

Formula (a4) represents, the genotype P of target SNP site father _gtduring for AA, P _1gtwith P _2gtequal value is A; The genotype P of target SNP site father _gtduring for BB, P _1gtwith P _2gtequal value is B;

Formula (a5) represents, asks R _gtwith P _1gtdifference set as M _1gtvalue.By R _gt(being made up of two equipotential types) removes P _1gtequipotential type remaining after the equipotential type represented is appointed as M _1gtvalue.

Formula (6) represents, and asks M _gtwith M _1gtdifference set as M _2gtvalue.By M _gt(being made up of two equipotential types) removes M _1gtequipotential type remaining after the equipotential type represented is as M _2gtvalue.In formula (6), M _gtvalue is AB.

Think under this situation, p1 and m1 is acquired haplotype, all non-acquired haplotype of p2 and m2.

To define certain site be the prerequisite in the effective tagSNP site of mother is be AB in the genotype of this site mother, and the genotype of father is AA or BB.In one embodiment, the genotype of father is AA, and propositus's genotype is AB, so P _1gtwith P _2gtequal value is A, R _gtwith M _gtvalue is AB; M _1gt=R _gt-P _1gt=AB-A=B; M _2gt=M _gt-M _1gt=AB-B=A.In another embodiment, the genotype of father is BB, and propositus's genotype is BB, so P _1gtwith P _2gtequal value is B, R _gtvalue is BB; M _1gt=R _gt-P _1gt=BB-B=B; M _2gt=M _gt-M _1gt=AB-B=A.

2) between propositus and candidate embryo, family's genealogical relationship is elder brother, when target gene is on X chromosome.

Mother effective tagSNP site computing formula is as follows:

$P_{1gt}=\left\{\begin{array}{c}A\left(P_{gt}=AA\right)\\ B\left(P_{gt}=BB\right)\end{array}\right.---\left(b4\right)$

$M_{1gt}=\left\{\begin{array}{c}A\left(R_{gt}=AA\right)\\ B\left(R_{gt}=BB\right)\end{array}\right.---\left(b5\right)$

M _2gt＝M _gt-M _1gt(6)

Formula (b4) represents, the genotype P of target SNP site father _gtduring for AA, P _1gtvalue is A; The genotype P of target SNP site father _gtduring for BB, P _1gtvalue is B;

Formula (b5) represents, the genotype R of target SNP site propositus _gtduring for AA, M _1gtvalue is A; The genotype R of target SNP site propositus _gtduring for BB, M _1gtvalue is B;

Formula (6) implication is the same.

Think under this situation, m1 is acquired haplotype, and p2 does not exist, and p1 and m2 is non-acquired haplotype.

To define certain site be the prerequisite in the effective tagSNP site of mother is genotype M this site mother _gtfor AB, because the male sex only has an X chromosome, so for the male sex, this SNP site genotype is just designated as homozygous AA or BB according to the allelic gene type in this site on its X chromosome, and namely the genotype of father is AA or BB.Because father is the male sex, only has X chromosome, therefore a P _2gtnon-existent.In one embodiment, the genotype P of father _gtfor AA, propositus genotype R _gtfor AA, so P _1gtvalue is A; M _1gt=A; M _2gt=M _gt-M _1gt=AB-A=B.In another embodiment, the genotype P of father _gtfor AA, propositus genotype R _gtfor BB, so P _1gtvalue is A; M _1gt=B; M _2gt=M _gt-M _1gt=AB-B=A.

3) between propositus and candidate embryo, family's genealogical relationship is elder sister, when target gene is on X chromosome.

Mother effective tagSNP site computing formula is as follows:

$P_{1gt}=\left\{\begin{array}{c}A\left(P_{gt}=AA\right)\\ B\left(P_{gt}=BB\right)\end{array}\right.---\left(b4\right)$

M _1gt＝R _gt-P _1gt(a5)

M _2gt＝M _gt-M _1gt(6)

Formula (b4), (a5), (6) implication are the same.

To define certain site be the prerequisite in the effective tagSNP site of mother is genotype M this site mother _gtfor AB, the genotype of father is AA or BB.Because father is the male sex, only has X chromosome, therefore a P _2gtnon-existent.In one embodiment, the genotype P of father _gtfor AA, propositus genotype R _gtfor AB, so P _1gt=A; M _1gt=R _gt-P _1gt=AB-A=B; M _2gt=M _gt-M _1gt=AB-B=A.In another embodiment, the genotype P of father _gtfor BB, propositus genotype R _gtfor BB, so P _1gtfor B; M _1gt=R _gt-P _1gt=BB-B=B; M _2gt=M _gt-M _1gt=AB-B=A.

Think under this situation, m1 and p1 is acquired haplotype, and p2 does not exist, and m2 is non-acquired haplotype.

4) between propositus and candidate embryo, family's genealogical relationship is grandfather, grandmother, aunt or uncle, when target gene is on autosome.

Father effective tagSNP site computing formula is as follows:

$M_{1gt}=M_{2gt}=\left\{\begin{array}{c}A\left(M_{gt}=AA\right)\\ B\left(M_{gt}=BB\right)\end{array}\right.---\left(1\right)$

P _1gt＝R _gt∩P _gt(R _gt≠AB)(b2)

P _2gt＝P _gt-P _1gt(3)

Formula (1), (3) represent that implication is the same;

Formula (b2) represents, asks R _gtwith P _gtcommon factor as P _1gtvalue.By R _gt(being made up of two equipotential types) and P _gtthe equipotential type that (being made up of two equipotential types) has is as P _1gtvalue.In formula, R _gtcan not value be AB.As R _gtvalue is AB, so P _1gtwith P _2gtwithout separating.

Think under this situation, p1 is acquired haplotype, and p2, m1 and m2 are non-acquired haplotype.

The effective tagSNP site of mother cannot calculate.

To define certain site be the prerequisite in the effective tagSNP site of father is genotype P this site father _gtfor AB, the genotype of mother is AA or BB.In one embodiment, the genotype M of mother _gtfor AA, propositus's genotype is AA, so M _1gtwith M _2gtequal value is A; P _1gtequipotential type identical with propositus's genotype (AA) in the genotype (AB) of father, i.e. A equipotential; P _2gtfor P _2gt=P _gt-P _1gt=AB-A=B.In another embodiment, the genotype of mother is BB, and propositus's genotype is AB, so M _1gtwith M _2gtequal value is B; Because the genotype of father is identical with the genotype of propositus, therefore P _1gtand P _2gtjust have no idea to determine.

5) between propositus and candidate embryo, family's genealogical relationship is granddad, grandmother, aunt, uncle, when target gene is on autosome.

Mother effective tagSNP site computing formula is as follows:

$P_{\lg t}=P_{2gt}=\left\{\begin{array}{c}A\left(P_{gt}=AA\right)\\ B\left(P_{gt}=BB\right)\end{array}\right.---\left(a4\right)$

M _1gt＝R _gt∩M _gt(R _gt≠AB)(c5)

M _2gt＝M _gt-M _1gt(6)

Formula (c5) represents, asks R _gtwith M _gtcommon factor as M _1gtvalue.By R _gt(being made up of two equipotential types) and M _gtthe equipotential type that (being made up of two equipotential types) has is as M _1gtvalue.In formula, R _gtcan not value be AB.As R _gtvalue is AB, so M _1gtwith M _2gtwithout separating.

Formula (a4), (6) implication are the same.

Think under this situation, m1 is acquired haplotype, and m2, p1 and p2 are non-acquired haplotype.

The effective tagSNP site of father cannot calculate.

To define certain site be the prerequisite in the effective tagSNP site of mother is genotype M this site mother _gtfor AB, the genotype of father is AA or BB.In one embodiment, the genotype of father is BB, and propositus's genotype is BB, so P _1gtwith P _2gtequal value is B, R _gtvalue is BB; M _1gtfor B; M _2gt=M _gt-M _1gt=AB-B=A.

6) between propositus and candidate embryo, family's genealogical relationship is granddad or uncle, when target gene is on X chromosome.

Mother effective tagSNP site computing formula is as follows:

$P_{1gt}=\left\{\begin{array}{c}A\left(P_{gt}=AA\right)\\ B\left(P_{gt}=BB\right)\end{array}\right.---\left(b4\right)$

$M_{1gt}=\left\{\begin{array}{c}A\left(R_{gt}=AA\right)\\ B\left(R_{gt}=BB\right)\end{array}\right.---\left(b5\right)$

M _2gt＝M _gt-M _1gt(6)

Formula (b4), (b5), (6) implication are the same.

Think under this situation, m1 is acquired haplotype, and p2 does not exist, and p1 and m2 is non-acquired haplotype.

To define certain site be the prerequisite in the effective tagSNP site of mother is genotype M this site mother _gtfor AB, the genotype of father is AA or BB.Because father is the male sex, only has X chromosome, therefore a P _2gtnon-existent.In one embodiment, the genotype P of father _gtfor BB, propositus genotype R _gtfor BB, so P _1gtfor B; M _1gtfor B; M _2gt=M _gt-M _1gt=AB-B=A.

7) between propositus and candidate embryo, family's genealogical relationship is grandmother or aunt, when target gene is on X chromosome.

Mother effective tagSNP site computing formula is as follows:

$P_{1gt}=\left\{\begin{array}{c}A\left(P_{gt}=AA\right)\\ B\left(P_{gt}=BB\right)\end{array}\right.---\left(b4\right)$

M _1gt＝R _gt∩M _gt(R _gt≠AB)(c5)

M _2gt＝M _gt-M _1gt(6)

Formula (a4), (c5), (6) implication are the same.

Think under this situation, m1 is acquired haplotype, and p2 does not exist, and p1 and m2 is non-acquired haplotype.

To define certain site be the prerequisite in the effective tagSNP site of mother is genotype M this site mother _gtfor AB, the genotype of father is AA or BB.Because father is the male sex, only has X chromosome, therefore a P _2gtnon-existent.In one embodiment, the genotype P of father _gtfor BB, propositus genotype R _gtfor BB, so P _1gtfor B; M _1gtfor B; M _2gt=M _gt-M _1gt=AB-B=A.

Be grandfather, grandmother, uncle, aunt for family's genealogical relationship between propositus and candidate embryo, when target gene is on X chromosome, according to existing science of heredity general knowledge, without the need to adopting device of the present invention to judge, only need simply to judge whether candidate embryo carries gene defect and whether phenotype is normal according to the sex of the phenotype of father, the mode of inheritance of single gene inheritance disease and embryo.

According to above formula, first we build according to the equipotential type of each site on parental chromosomes in the target SNP site combination of target gene the haplotype obtaining target gene parents.Such as, certain target gene be positioned in normal dyeing defines 10 target SNP site, and determine that its equipotential type on p1 is A, A, B, A, A, A, B, B, A and B respectively, so the formation of target gene haplotype p1 is AABAAABBAB.

Candidate embryo haplotype reconstruction and genetic origin determination module 300

Crucial equipotential: on the effective tagSNP of father or the effective tagSNP site of mother, has an equipotential type and other equipotential types are different, and this equipotential type is crucial equipotential.

Crucial SNP site: be selected from target SNP site, the target SNP site for candidate embryo comprises the target SNP site of crucial equipotential.

Not crucial SNP site: be selected from target SNP site, the target SNP site for candidate embryo does not comprise the target SNP site of crucial equipotential.

The introducing of crucial equipotential and crucial SNP site is the problem in order to avoid allele dropout (Alleledrop-out, ADO).Up to the present, the unicellular gene magnification of embryo still can not avoid the problem of ADO completely.ADO is because very few one of them the caused allele of test material does not increase, or amplification quantity not sufficient is to reach the level of detecting, and it can cause the erroneous judgement of PGD result, directly can cause the implantation of abnormal embryo in extreme circumstances, cause serious consequence.Such as, analyzing in the genotype results obtained, if the genotype in this site is for isozygotying, we can not affirm inherently isozygoty in this site, or there occurs ADO in amplification.Because being likely this site is be heterozygous originally, in amplification procedure, there occurs ADO, lost an equipotential, thus become false homozygous.In this case, the judgement of the haplotype of embryo being originated has a great impact.Therefore, introduce the concept of crucial equipotential and crucial SNP when we analyze: under normal circumstances, on autosome, on same tagSNP site, parents always have four equipotentials; On X chromosome, parents always have three equipotentials.Effective tagSNP site has an equipotential certainly and other equipotentials different, we are defined as this equipotential the crucial equipotential (KB) on this tagSNP site.In one embodiment, father is AB, and mother is AA, and so B is exactly crucial equipotential (KB=B).Therefore, if this site of embryo does not comprise crucial equipotential, this site is likely that ADO causes, and the confidence level that the embryo chromosome source in this site is distinguished is more weak, and the SNP in this site is defined as not crucial SNP (Non-keySNP); If same this site of embryo comprises crucial equipotential, the confidence level that the embryo chromosome source in this site is distinguished is comparatively strong, and the SNP in this site is defined as crucial SNP (KeySNP).

In the embodiment of the present invention, the haplotype of target gene on two chromosomes of candidate embryo is denoted as E1 and E2 respectively, and wherein E1 refers to the haplotype from father, and E2 refers to the haplotype from mother.

The haplotype of target gene on two chromosomes of candidate embryo is combined by crucial SNP site and forms.

The target SNP site of candidate embryo comprises two equipotentials, lays respectively on male parent chromosome and maternal chromosome, and the parental source of the equipotential on male parent chromosome is P1, P2, NA or NB, and the parental source of the equipotential on maternal chromosome is M1, M2 or NA.

In one embodiment, adopt whether following formula determination target SNP site is crucial SNP site, and obtain crucial SNP site two chromosomal parental sources on candidate embryo.

Alphabetical implication in formula:

E _1O: the parental source of target SNP site on the male parent chromosome referring to candidate embryo, its value can be P1, P2, NA or NB

E _2O: the parental source of target SNP site on the maternal chromosome referring to candidate embryo, its value can be M1, M2 or NA

P1: represent relevant to candidate father embryo, and have genetic association with children propositus or paternal propositus

P2: represent relevant to candidate father embryo, and with children propositus or paternal propositus without genetic association

M1: represent relevant to candidate mother embryo, and have genetic association with children propositus or maternal propositus

M2: represent relevant to candidate mother embryo, and with children propositus or maternal propositus without genetic association

NA: represent and exist, but cannot to judge or without the need to determining

NB: represent and do not exist

E _1gt: the equipotential type referring to target SNP site in haplotype E1 on autosome or X chromosome, its value can be A or B; When candidate embryo is the male sex, E _1gtdo not exist.

E _2gt: the equipotential type referring to target SNP site in haplotype E2 on autosome or X chromosome, its value can be A or B;

E1: refer to the target gene haplotype of candidate embryo from father; E2: refer to the target gene haplotype of candidate embryo from mother; When candidate embryo is the male sex, E1 does not exist.

E _gt: the genotype referring to candidate embryo, its value is AA, BB or AB, and its value derives from message unit;

KB: refer to crucial equipotential, its value can be A or B;

E _r: judge whether target SNP site is crucial SNP site, and value can be KeySNP or Non-KeySNP.

1) when family's genealogical relationship of propositus and candidate embryo is elder brother or elder sister, when target gene is on autosome.

Father effective tagSNP site computing formula is as follows:

E _2O＝“NA”(a7)

$E_{1gt}=\left\{\begin{array}{c}B\left(M_{gt}=AA,E_{gt}=BB\right)\\ A\left(M_{gt}=BB,E_{gt}=AA\right)\\ E_{gt}-M_{\lg t}\end{array}\right.---\left(a8\right)$

$KB=\left\{\begin{array}{c}A\left(M_{gt}=BB\right)\\ B\left(M_{gt}=AA\right)\end{array}\right.---\left(a10\right)$

Formula (a7) represents: E _2Ocannot to judge or without the need to determining.

Formula (a8) represents: M _gtfor AA and E _gtduring for BB (there occurs ADO when this situation is the SNP site detection of candidate embryo, thus cause the loss of the base testing result from mother), E _1gtvalue is B; M _gtfor BB and E _gtfor (there occurs ADO when this situation is also the SNP site detection of candidate embryo, thus cause the loss of the base testing result from mother) during AA, E _1gtvalue is A; In other situations, ask E _gtwith M _1gtdifference set as E _1gtvalue.

Formula (a9) represents: work as E _1gtwith P _1gtwhen value is equal, E _1Ovalue is P1; Work as E _1gtwith P _2gtwhen value is equal, E _1Ovalue is P2.

Formula (a10) represents: work as M _gtduring for BB, KB value is A; Work as M _gtduring for AA, KB value is B;

Formula (a11) represents: work as E _1gttime equal with KB value, E _rvalue is KeySNP; Work as E _1gttime unequal with KB value, E _rvalue is Non-KeySNP.

Think under this situation, p1 and m1 is acquired haplotype, p2 and m2 is non-acquired haplotype.Because the equipotential type of this SNP site on p1 and p2 can be distinguished, thus can determine which equipotential type and propositus have genetic association.Due to the equipotential type undistinguishable of this SNP site on m1 and m2, thus cannot determine which equipotential type and propositus have genetic association.Therefore, the parental source that this SNP site candidate father embryo is correlated with can only be determined under this situation, and the parental source that candidate mother embryo is correlated with cannot be determined.

Mother effective tagSNP site computing formula is as follows:

E _1O＝“NA”(b9)

$E_{2gt}=\left\{\begin{array}{c}B\left(P_{gt}=AA,E_{gt}=BB\right)\\ A\left(P_{gt}=BB,E_{gt}=AA\right)\\ E_{gt}-P_{1gt}\end{array}\right.---\left(a12\right)$

$KB=\left\{\begin{array}{c}A\left(P_{gt}=BB\right)\\ B\left(P_{gt}=AA\right)\end{array}\right.---\left(b10\right)$

Formula (b9) represents: E _1Ocannot to judge or without the need to determining.

Formula (a12) represents: P _gtfor AA and E _gtfor (there occurs ADO when this situation is the SNP site detection of candidate embryo, thus cause the loss of the base testing result from father) during BB, E _2gtvalue is B; P _gtfor BB and E _gtfor (there occurs ADO when this situation is the SNP site detection of candidate embryo, thus cause the loss of the base testing result from father) during AA, E _2gtvalue is A; In other situations, ask E _gtwith P _1gtdifference set as E _2gtvalue.

Formula (b7) represents: work as E _2gtwith M _1gtwhen value is equal, E ₂₀value is M1; Work as E _2gtwith M _2gtwhen value is equal, E _2Ovalue is M2.

Formula (b10) represents: work as P _gtduring for BB, KB value is A; Work as P _gtduring for AA, KB value is B;

Formula (b11) represents: work as E _2gttime equal with KB value, E _rvalue is KeySNP; Work as E _2gttime unequal with KB value, E _rvalue is Non-KeySNP.

Think under this situation, p1 and m1 is acquired haplotype, p2 and m2 is non-acquired haplotype.Because the equipotential type of this SNP site on m1 and m2 can be distinguished, thus can determine which and propositus have genetic association.Due to the equipotential type undistinguishable of this SNP site on p1 and p2, thus cannot determine which and propositus have genetic association.Therefore, the parental source that this SNP site candidate mother embryo is correlated with can only be determined under this situation, and the parental source that candidate father embryo is correlated with cannot be determined.

2) be man when family's genealogical relationship of propositus and candidate embryo is the sex of elder brother or elder sister and embryo, when target gene is on X chromosome.

Mother effective tagSNP site computing formula is as follows:

E _1O＝“NB”(d9)

$E_{2gt}=\left\{\begin{array}{c}A\left(E_{gt}=AA\right)\\ B\left(E_{gt}=BB\right)\end{array}\right.---\left(b12\right)$

$KB=\left\{\begin{array}{c}A\left(P_{gt}=BB\right)\\ B\left(P_{gt}=AA\right)\end{array}\right.---\left(b10\right)$

Because embryo is the male sex, only has X chromosome, therefore an E _1gtnon-existent.

Formula (d9) represents: E _1Odo not exist.

Formula (b12) represents: E _gtduring for AA, E _2gtvalue is A; E _gtduring for BB, E _2gtvalue is B;

Formula (b7) represents: work as E _2gtwith M _1gtwhen value is equal, E _2Ovalue is M1; Work as E _2gtwith M _2gtwhen value is equal, E _2Ovalue is M2.

Think under this situation, when propositus family's genealogical relationship is elder brother, m1 is acquired haplotype, and p2 does not exist, and p1 and m2 is non-acquired haplotype; When propositus family's genealogical relationship is elder sister, m1 and p1 is acquired haplotype, and p2 does not exist, and m2 is non-acquired haplotype.Because the equipotential type of this SNP site on m1 and m2 can be distinguished, thus can determine which and propositus have genetic association.Because embryo is the male sex, do not have heredity from the X chromosome of father, therefore only need determine the parental source that this SNP site candidate mother embryo is correlated with, the information that this SNP site candidate father embryo is correlated with does not exist.

3) sex being elder brother or elder sister and embryo when family's genealogical relationship of propositus and candidate embryo is female, when target gene is on X chromosome.

Mother effective tagSNP site computing formula is as follows:

E _1gt＝P _1gt(b8)

E _1O＝“P1”(c9)

$E_{2gt}=\left\{\begin{array}{c}B\left(P_{gt}=AA,E_{gt}=BB\right)\\ A\left(P_{gt}=BB,E_{gt}=AA\right)\\ E_{gt}-P_{1gt}\end{array}\right.---\left(a12\right)$

$KB=\left\{\begin{array}{c}A\left(P_{gt}=BB\right)\\ B\left(P_{gt}=AA\right)\end{array}\right.---\left(b10\right)$

Formula (b8) represents, E _1gtwith P _1gtvalue is equal

Formula (c9) represents, E _1Ovalue is P1

Think under this situation, when propositus family's genealogical relationship is elder brother, m1 is acquired haplotype, and p2 does not exist, and p1 and m2 is non-acquired haplotype; When propositus family's genealogical relationship is elder sister, m1 and p1 is acquired haplotype, and p2 does not exist, and m2 is non-acquired haplotype.Because the equipotential type of this SNP site on m1 and m2 can be distinguished, thus can determine which and propositus have genetic association.Because SNP site is positioned at X chromosome and embryo is women, and candidate father embryo only has an X chromosome, the X chromosome that therefore the inevitable hereditary father of candidate embryo is unique.Therefore, the parental source that this SNP site candidate mother embryo is correlated with and the parental source that this SNP site candidate father embryo is correlated with can be determined in this case.

4) when family's genealogical relationship of propositus and candidate embryo is grandfather, grandmother, aunt or uncle, when target gene is on autosome.

Father effective tagSNP site computing formula is as follows:

E _2O＝“NA”(a7)

$E_{1gt}=\left\{\begin{array}{c}B\left(M_{gt}=AA,E_{gt}=BB\right)\\ A\left(M_{gt}=BB,E_{gt}=AA\right)\\ E_{gt}-M_{\lg t}\end{array}\right.---\left(a8\right)$

$KB=\left\{\begin{array}{c}A\left(M_{gt}=BB\right)\\ B\left(M_{gt}=AA\right)\end{array}\right.---\left(a10\right)$

Formula (a7), (a8), (a9) (a10) (a11) implication are the same.

Think under this situation, p1 is acquired haplotype, and p2, m1 and m2 are non-acquired haplotype.Because the equipotential type of this SNP site on p1 and p2 can be distinguished, thus can determine which and propositus have genetic association.Because propositus is paternal propositus, with candidate mother embryo without genetic association, lower of this situation need determine parental source relevant to candidate father embryo in this SNP site, and parental source relevant to candidate mother embryo in this SNP site is without the need to determining.

The effective tagSNP site of mother cannot calculate.

Think under this situation, p1 is acquired haplotype, and p2, m1 and m2 are non-acquired haplotype.Due to the equipotential type undistinguishable of this SNP site on p1 and p2, thus can not determine which and propositus have genetic association.Parental source relevant to candidate father embryo in this SNP site is can not determine under this situation.

5) when family's genealogical relationship of propositus and candidate embryo is granddad, grandmother, aunt, uncle, when target gene is on autosome.

Mother effective tagSNP site computing formula is as follows:

E _1O＝“NA”(b9)

$E_{2gt}=\left\{\begin{array}{c}B\left(P_{gt}=AA,E_{gt}=BB\right)\\ A\left(P_{gt}=BB,E_{gt}=AA\right)\\ E_{gt}-P_{1gt}\end{array}\right.---\left(a12\right)$

$KB=\left\{\begin{array}{c}A\left(P_{gt}=BB\right)\\ B\left(P_{gt}=AA\right)\end{array}\right.---\left(b10\right)$

Formula (b9), (a12), (b7) (b10) (b11) implication are the same.

Think under this situation, m1 is acquired haplotype, and m2, p1 and p2 are non-acquired haplotype.Because the equipotential type of this SNP site on m1 and m2 can be distinguished, thus can determine which and propositus have genetic association.Because propositus is maternal propositus, with candidate father embryo without genetic association, lower of this situation need determine parental source relevant to candidate mother embryo in this SNP site, and parental source relevant to candidate father embryo in this SNP site is without the need to determining.

The effective tagSNP site of father cannot calculate.

Think under this situation, m1 is acquired haplotype, and m2, p1 and p2 are non-acquired haplotype.Due to the equipotential type undistinguishable of this SNP site on m1 and m2, thus can not determine which and propositus have genetic association.Parental source relevant to candidate mother embryo in this SNP site is can not determine under this situation.

6) when family's genealogical relationship of propositus and candidate embryo be granddad, grandmother, aunt, uncle and embryo sex be man, when target gene is on X chromosome.

Mother effective tagSNP site computing formula is as follows:

E _1O＝“NB”(d9)

$E_{2gt}=\left\{\begin{array}{c}A\left(E_{gt}=AA\right)\\ B\left(E_{gt}=BB\right)\end{array}\right.---\left(b12\right)$

$KB=\left\{\begin{array}{c}A\left(P_{gt}=BB\right)\\ B\left(P_{gt}=AA\right)\end{array}\right.---\left(b10\right)$

Formula (d9), (b12), (b7) (b10) (b11) implication are the same.

Think under this situation, m1 is acquired haplotype, and p2 does not exist, and p1 and m2 is non-acquired haplotype.Because the equipotential type of this SNP site on m1 and m2 can be distinguished, thus can determine which and propositus have genetic association.Because embryo is the male sex, do not have heredity from the X chromosome of father, therefore only need determine the parental source that this SNP site candidate mother embryo is correlated with, the information that this SNP site candidate father embryo is correlated with does not exist.

7) when family's genealogical relationship of propositus and candidate embryo be granddad, grandmother, aunt, uncle and embryo sex be female, when target gene is on X chromosome.

Mother effective tagSNP site computing formula is as follows:

E _1gt＝P _1gt(b8)

E _1O＝“NA”(b9)

$E_{2gt}=\left\{\begin{array}{c}B\left(P_{gt}=AA,E_{gt}=BB\right)\\ A\left(P_{gt}=BB,E_{gt}=AA\right)\\ E_{gt}-P_{1gt}\end{array}\right.---\left(a12\right)$

$KB=\left\{\begin{array}{c}A\left(P_{gt}=BB\right)\\ B\left(P_{gt}=AA\right)\end{array}\right.---\left(b10\right)$

Formula (b8), (b9), (a12) (b7) (b10) (b11) implication are the same.

Think under this situation, m1 is acquired haplotype, and p2 does not exist, and p1 and m2 is non-acquired haplotype.Because the equipotential type of this SNP site on m1 and m2 can be distinguished, thus can determine which and propositus have genetic association.Because propositus is maternal propositus, with candidate father embryo without genetic association, therefore only need determine the parental source that this SNP site candidate mother embryo is correlated with, parental source relevant to candidate father embryo in this SNP site is without the need to determining.

Candidate embryo haplotype parental source determination module 400

In a specific embodiment, as shown in Figure 5, candidate embryo haplotype parental source determination module 400 comprises further:

Candidate embryo haplotype parental source statistical module 410, be connected with candidate embryo haplotype reconstruction and genetic origin determination module 300 and amphiphilic monomer type creation module 200, for the crucial SNP site combination corresponding according to target gene candidate embryo haplotype, and each crucial SNP site two chromosomal parental sources, in statistics target gene candidate embryo haplotype, the crucial SNP site sum that all kinds of crucial SNP site two chromosomal parental sources are corresponding.

Concrete, crucial SNP site two chromosomal parental sources have four classes: " P1 " " P2 " " M1 " " M2 ".According to the chromosomal parental source result of each crucial SNP site two obtained in module 300, add up in target gene candidate embryo haplotype respectively, the crucial SNP site sum P of crucial SNP site two chromosomal parental source difference corresponding " P1 " " P2 " " M1 " " M2 " _1N, P _2N, M _1N, M _2N.

Candidate embryo haplotype parental source analysis module 420, be connected with candidate embryo haplotype parental source statistical module 410, for the crucial SNP site sum according to all kinds of parental source of target gene candidate embryo haplotype, determine candidate embryo haplotype parental source.

Preferred further, when described candidate embryo haplotype parental source statistical module 410 is also 0 for the crucial SNP site sum corresponding when described all kinds of crucial SNP site two chromosomal parental sources, expand target area in amphiphilic monomer type creation module 200 to increase target SNP site number, until crucial SNP site sum corresponding to described all kinds of crucial SNP site two chromosomal parental sources has at least one not to be 0.

In one embodiment of the present of invention, adopt the parental source of following methods analyst determination target gene candidate embryo haplotype:

Symbol implication:

P _1N: in target gene candidate embryo haplotype, the parental source type of crucial SNP site is the crucial SNP site sum of " P1 ";

P _2N: in target gene candidate embryo haplotype, the parental source type of crucial SNP site is the crucial SNP site sum of " P2 ";

M _1N: in target gene candidate embryo haplotype, the parental source type of crucial SNP site is the crucial SNP site sum of " M1 ";

M _2N: in target gene candidate embryo haplotype, the parental source type of crucial SNP site is the crucial SNP site sum of " M2 ";

1) P _1N>0, P _2N=0, then candidate embryo haplotype E1 parental source is P1

2) P _1N=0, P _2N>0, then candidate embryo haplotype E1 parental source is P2

3) P _1N=0, P _2N=0, as each crucial SNP site E _1Owhen being NA, then the parental source of candidate embryo haplotype E1 cannot judge or without the need to determining, be NA; As each crucial SNP site E _1Owhen being NB, then the parental source of candidate embryo haplotype E1 does not exist, and is NB

4) P _1N>0, P _2N>0, when there is not crucial SNP site in the region at target gene place, the parental source from the nearest crucial SNP site in upstream of target gene and the crucial SNP site in nearest downstream is selected to add up: if the statistics in these two sites is P _1N=2P _2N=0, then candidate embryo haplotype E1 parental source is P1; If statistics is P _2N=2P _1N=0, then candidate embryo haplotype E1 parental source is P2; If statistics is P _1N=1, and P _2N=1, then candidate embryo haplotype E ₁source is P1/P2.When there is crucial SNP site in the region at target gene place, the parental source of the crucial SNP site in the region shared by target gene is added up: if the statistics in these sites is P _1N>0P _2N=0, then candidate embryo haplotype E1 parental source is P1; Statistics is P _1N=0P _2N>0, then candidate embryo haplotype E1 parental source is P2; Statistics is P _1N=0P _2N=0, then the parental source of candidate embryo haplotype E1 is NA or NB; Statistics is P _1N>0P _2N>0, then candidate embryo haplotype E ₁parental source is P1/P2

5) M _1N>0, M _2N=0, then candidate embryo haplotype E2 parental source is M1

6) M _1N=0, M _2N=0, then the parental source of candidate embryo haplotype E2 cannot judge, is NA

7) M _1N=0, M _2N>0, then candidate embryo haplotype E2 parental source is M2

8) M _1N>0, M _2N>0, when there is not crucial SNP site in the region at target gene place, the parental source from the nearest crucial SNP site in upstream of target gene and the crucial SNP site in nearest downstream is selected to add up: if the statistics in these two sites is M _1N=2M _2N=0, then candidate embryo haplotype E2 parental source is M1; If statistics is M _2N=2M _1N=0, then candidate embryo haplotype E2 parental source is M2; If statistics is M _1N=1, and M _2N=1, then candidate embryo haplotype E2 originates as M1/M2; When there is crucial SNP site in the region at target gene place, the parental source of the crucial SNP site in the region shared by target gene is added up: if the statistics in these sites is M _1N>0M _2N=0, then candidate embryo haplotype E2 parental source is M1, and statistics is M _1N=0M _2N>0, then candidate embryo haplotype E2 parental source is M2, and statistics is M _1N=0M _2N=0, then the parental source of candidate embryo haplotype E2 cannot judge, is NA, and statistics is M _1N>0M _2N>0, then candidate embryo haplotype E2 originates as M1/M2

Whether candidate embryo carries gene defect judge module 500

Judge whether embryo carries gene defect.When result is understood, need simultaneously to judge whether embryo carries gene defect with reference to the parental source of two chromosomal haplotypes of family's genealogical relationship of the mode of inheritance of hereditary disease, propositus and candidate embryo, father and mother's phenotype and candidate embryo.

Further, module 500 also can be used for judging that whether the phenotype of candidate embryo is normal.

In an embodiment of the present invention, utilize table 1 to contrast and judge whether candidate embryo carries gene defect and judge that whether the phenotype of candidate embryo is normal.

Table 1.PGD single-gene result is understood

The method for expressing of candidate embryo haplotype parental source is: " E1 parental source, E2 parental source ".Such as " P1, M1 " represents E1 parental source be P1, E2 parental source is M1.

NA represents and cannot to judge or without the need to determining parental source, NB represents not exist.

Candidate's embryo chromosome abnormality juding module 600

The region of chromosome abnormality is had specifically to refer to: the SNP site B gene frequency band in this region is compared normal phenotype and changed.

Situation of change comprises following two kinds:

3) in the SNP site B gene frequency figure in this region, the number of band increases, then think that chromosome number increases;

4) in the SNP site B gene frequency figure in this region, the decreased number of band, then think that chromosome number reduces.

B gene frequency figure is: the frequency distribution schematic diagram of the Type B equipotential SNP detected.In the present invention, base A and base T is defined as A type equipotential, and base C and bases G are defined as Type B equipotential.

The cytogenetics software package (such as, KaryoStudio or GenomeStudio of illumina) that this module can utilize any one SNP chip supporting has come.

Candidate embryo HLA distribution type analysis module 700

The HLA being carried out PIE by PGD is detected, and can select embryo's row transplantation of mating with the patient compatriot HLA needing to transplant candidate stem cell.Transplant successful key and be that whether the HLA type between donor with receptor is consistent.When analyzing, the detection whether carrying out HLA distribution type can be selected according to demand.

The gene that HLA is relevant comprises: HLA-A, HLA-B, HLA-C, HLA-DPB1, HLA-DQB1 and HLA-DRB1.

In one embodiment, analyze HLA be correlated with 6 genes embryo's haplotype source.These 6 genes and position thereof be respectively: HLA-A (chr6:29,910,247-29,913,661), HLA-B (chr6:31,321,649-31,324,989), HLA-C (chr6:31,236,526-31,239,913), HLA-DPB1 (chr6:33,043,703-33,057,473), HLA-DQB1 (chr6:32,627,241-32,634,466) and HLA-DRB1 (32,546,546-32,557,613).Therefore, when analyzing, we select to comprise this gene interval chr6:29 of 6,000,000-33,500,000.Under normal circumstances, propositus is P1, M1 at the haplotype parental source of this section, if if therefore embryo be P1, M1 at the haplotype parental source of this section, illustrate and to mate with the HLA of patient.Otherwise, do not mate.

Sample Quality Control unit 800

The threshold value of the recall rate of the SNP site of candidate embryo sample mother and the SNP site of propositus can rule of thumb set, the recall rate that such as can set the SNP site of parents and propositus reports an error lower than 70%, and the recall rate of the SNP site of embryo's sample reports an error lower than 50%.

Judge that whether the sex input of candidate embryo sample parents' sex and propositus is wrong to refer to: whether there is any discrepancy for the candidate embryo sample father and mother of inspection input, the sex of propositus and actual sample, and there is any discrepancy then reports an error.

As the highly preferred embodiment of the present invention, as shown in Figure 6, all modules in prostatitis are comprised.

Following examples, with the test-tube baby of three beta Thalassemia familys alternatively embryo, utilize the science of heredity pick-up unit before Embryonic limb bud cell of the present invention to screen these three candidate embryos.

β-thalassemia (be called for short β ground poor) is 0.67% at the south China incidence of disease, and the incidence of disease of south China, southwest is more, and patient, except seeing Han nationality, also sees strong, multitude, seedling, returns and Yao Deng11Ge ethnic group.β ground is poor very big to human health risk, poor patient especially heavyly, and prognosis seriously, second brings heavy burden to family, society first.Science of heredity pick-up unit before the present embodiment utilizes Embryonic limb bud cell of the present invention is detected by heredity before Embryonic limb bud cell can avoid the appearance of this type of infant, selects the embryo identical with infant HLA distribution type pass through the to give a birth bleeding of the umbilicus of baby or marrow can treat existing infant simultaneously.In present case, the normal Mr. and Mrs of certain phenotype have one and suffer from the daughter of beta Thalassemia, and these Mr. and Mrs want to obtain a healthy baby by the means of third generation test-tube baby, and wish that the bleeding of the umbilicus of this baby or marrow are to treat existing daughter.In present case, Disease-causing gene is HBB, is positioned on autosome, and mode of inheritance is autosomal recessive, and propositus is the daughter of these father and mother.3 well-developed embryos are obtained by third generation vitro techniques.Therefore, the haplotype source of the HBB gene of these three embryos of following Water demand and HLA distribution type related gene, thus obtain the healthy and fetal tissues of mating with daughter HLA, transplant.

First, chip detection is carried out to father and mother and propositus's sample.Basic step comprise extract DNA, DNA cloning, fragmentation, precipitation, resuspended, with chip hybridization, chip extension, dyeing and wrap SNP site information, the SNP site information of candidate embryo sample mother, the SNP site information of propositus, the SNP site information of candidate embryo sample that quilt, chip scanning etc. obtain the candidate embryo sample father that message unit 100 needs.

The SNP site information of candidate embryo sample father, the SNP site information of candidate embryo sample mother, candidate embryo sample father and mother's phenotypic information, the SNP site information of propositus, the SNP site information of candidate embryo sample is imported, the individual information of the individual information of propositus, candidate embryo sample father and mother's phenotypic information, propositus in message unit 100.

In this example, candidate embryo sample father and mother's phenotype are: father is normal, mother is normal.The individual information of propositus is: family's genealogical relationship of propositus and candidate embryo is elder sister.Present case examination disease is thalassemia, and therefore setting HBB is target gene, to should the mode of inheritance information of single gene inheritance disease of target gene be: autosomal recessive.

Utilize the science of heredity pick-up unit before Embryonic limb bud cell of the present invention, need suppose that amphilepsis on this target area all cross exchanged does not occur the respective haplotype of propositus to when.

Whether carry gene defect judge module 500 through amphiphilic monomer type creation module 200, candidate embryo haplotype reconstruction module 300, candidate embryo haplotype parental source determination module 400, candidate embryo successively, first establish: form the target SNP site combination of target gene HBB amphiphilic monomer type, two chromosomal equipotential types of each target SNP site parents; Then determine: the crucial SNP site combination forming target gene HBB candidate embryo haplotype, and each crucial SNP site two chromosomal parental sources in candidate embryo; Determine again: the parental source of target gene HBB candidate embryo haplotype; And then judge: whether candidate embryo carries gene defect.Final acquisition following table testing result, namely in 3 embryos, embryo 1 is all normal with the phenotype of embryo 3, and embryo 1 causes a disease (table 2).

The PGD result of table 23 embryo

Candidate's embryo chromosome abnormality juding module 600 is utilized to analyze 3 embryo chromosomes further all normal.

Utilize candidate embryo HLA distribution type analysis module 700 further, the haplotype parental source analyzed on gene region that acquisition 3 embryos are correlated with in HLA distribution type is respectively P1, M1, P1, M1 and P2, M2.

Phenotype due to embryo 1 is normal and mate with our propositus HLA, advises transplanting.

Above embodiment is in order to embodiment disclosed by the invention is described, can not be interpreted as limitation of the present invention.In addition, various amendment listed herein and invention in method, device change, be apparent concerning those skilled in the art without departing from the scope and spirit in the present invention.Although in conjunction with multiple concrete preferred embodiment of the present invention to invention has been concrete description, should be appreciated that the present invention should not be only limitted to these specific embodiments.In fact, variously as above invention is obtained concerning apparent amendment those skilled in the art and all should comprise within the scope of the invention.

Claims (16)

1. the science of heredity pick-up unit before Embryonic limb bud cell, is characterized in that, comprising:

Message unit 100, for obtaining the SNP site information of candidate embryo sample father, the SNP site information of candidate embryo sample mother, candidate embryo sample father and mother's phenotypic information, the SNP site information of propositus, the SNP site information of candidate embryo sample, the individual information of propositus, and store single gene inheritance disease information;

Amphiphilic monomer type creation module 200, be connected with SNP site message unit 100, the individual information for the SNP site information of the SNP site information of the SNP site information according to candidate embryo sample father, candidate embryo sample mother, propositus, single gene inheritance disease information and propositus is established: the target SNP site combination of the target gene that amphiphilic monomer type is corresponding; Two chromosomal equipotential types of each target SNP site parents thus build amphiphilic monomer type;

Candidate embryo haplotype reconstruction module 300, be connected with amphiphilic monomer type creation module 200 and message unit 100, determine for the SNP site information of the SNP site information of the SNP site information according to candidate embryo sample father, candidate embryo sample mother, candidate embryo sample, the target SNP site combination of target gene that amphiphilic monomer type is corresponding, two chromosomal equipotential types of each target SNP site parents: the crucial SNP site combination of the target gene that candidate embryo haplotype is corresponding, each crucial SNP site two chromosomal parental sources in candidate embryo;

Candidate embryo haplotype parental source determination module 400, be connected with candidate embryo haplotype reconstruction module 300, for the crucial SNP site combination according to target gene corresponding to candidate embryo haplotype, and each crucial SNP site two chromosomal parental sources are determined: the parental source of target gene candidate embryo haplotype;

Whether candidate embryo carries gene defect judge module 500, be connected with candidate embryo haplotype parental source determination module 400 and message unit 100, parental source, judgement for two chromosomal haplotypes of the candidate embryo according to single gene inheritance disease information, propositus's individual information, candidate embryo sample father and mother's phenotypic information, target gene: whether candidate embryo carries the gene defect in parents.

2. the science of heredity pick-up unit before Embryonic limb bud cell as claimed in claim 1, is characterized in that, in described message unit 100: each SNP site information at least comprises: the chromosome position of SNP, base composition and genotype information;

Single gene inheritance disease information at least comprises: the mode of inheritance information of the gene information that single gene inheritance disease is corresponding, single gene inheritance disease;

The individual information of propositus at least comprises: family's genealogical relationship of propositus and candidate embryo.

3. the science of heredity pick-up unit before Embryonic limb bud cell as claimed in claim 1, it is characterized in that, described amphiphilic monomer type creation module 200 comprises:

The target SNP site combination determination module 210 of target gene, be connected with message unit 100, for for target gene target setting region, determine according to the SNP site information of candidate embryo sample father in target area, the SNP site information of candidate embryo sample mother, single gene inheritance disease information: the target SNP site combination of the target gene that target gene amphiphilic monomer type is corresponding;

Two chromosomal equipotential determination type module 220 of each target SNP site parents, determination module 210 is combined and SNP site message unit 100 is connected with the target SNP site of target gene, for utilizing the SNP site information of candidate embryo sample father, the SNP site information of candidate embryo sample mother, the SNP site information of propositus, family's genealogical relationship between propositus and candidate embryo, the target SNP site combined information of target gene, determine: in the target SNP site combination of described target gene, the equipotential type of each target SNP site on two chromosomes of candidate embryo parents, thus build amphiphilic monomer type.

4. the science of heredity pick-up unit before Embryonic limb bud cell as claimed in claim 1, it is characterized in that, in described amphiphilic monomer type creation module 200, two chromosomal equipotential types of each target SNP site parents utilize following formula to determine:

1) between propositus and candidate embryo, family's genealogical relationship is elder brother or elder sister, when target gene is on autosome, as follows to father effective tagSNP site computing formula:

$M_{1gt}=M_{2gt}=\left\{\begin{array}{c}A(M_{gt}=AA)\\ B(M_{gt}=BB)\end{array}\right.---\left(1\right)$

P _1gt＝R _gt-M _1gt(a2)

P _2gt＝P _gt-P _1gt(3)

Mother effective tagSNP site computing formula is as follows:

$P_{1gt}=P_{2gt}=\left\{\begin{array}{c}A(P_{gt}=AA)\\ B(P_{gt}=BB)\end{array}\right.---\left(a4\right)$

M _1gt＝R _gt-P _1gt(a5)

M _2gt＝M _gt-M _1gt(6)

2) between propositus and candidate embryo, family's genealogical relationship is elder brother, and when target gene is on X chromosome, mother effective tagSNP site computing formula is as follows:

$P_{1gt}=\left\{\begin{array}{c}A(P_{gt}=AA)\\ B(P_{gt}=BB)\end{array}\right.---\left(b4\right)$

$M_{1gt}=\left\{\begin{array}{c}A(R_{gt}=AA)\\ B(R_{gt}=BB)\end{array}\right.---\left(b5\right)$

M _2gt＝M _gt-M _1gt(6)

3) between propositus and candidate embryo, family's genealogical relationship is elder sister, and when target gene is on X chromosome, mother effective tagSNP site computing formula is as follows:

$P_{1gt}=\left\{\begin{array}{c}A(P_{gt}=AA)\\ B(P_{gt}=BB)\end{array}\right.---\left(b4\right)$

M _1gt＝R _gt-P _1gt(a5)

M _2gt＝M _gt-M _1gt(6)

4) between propositus and candidate embryo, family's genealogical relationship is grandfather, grandmother, aunt or uncle, when target gene is on autosome

Father effective tagSNP site computing formula is as follows:

$M_{1gt}=M_{2gt}=\left\{\begin{array}{c}A(M_{gt}=AA)\\ B(M_{gt}=BB)\end{array}\right.---\left(1\right)$

P _1gt＝R _gt∩P _gt(R _gt≠AB)(b2)

P _2gt＝P _gt-P _1gt(3)

5) between propositus and candidate embryo, family's genealogical relationship is granddad, grandmother, aunt, uncle, when target gene is on autosome

Mother effective tagSNP site computing formula is as follows:

$P_{1gt}=P_{2gt}=\left\{\begin{array}{c}A(P_{gt}=AA)\\ B(P_{gt}=BB)\end{array}\right.---\left(a4\right)$

M _1gt＝R _gt∩M _gt(R _gt≠AB)(c5)

M _2gt＝M _gt-M _1gt(6)

6) between propositus and candidate embryo, family's genealogical relationship is granddad or uncle, and when target gene is on X chromosome, mother effective tagSNP site computing formula is as follows:

$P_{1gt}=\left\{\begin{array}{c}A(P_{gt}=AA)\\ B(P_{gt}=BB)\end{array}\right.---\left(b4\right)$

$M_{1gt}=\left\{\begin{array}{c}A(R_{gt}=AA)\\ B(R_{gt}=BB)\end{array}\right.---\left(b5\right)$

M _2gt＝M _gt-M _1gt(6)

7) between propositus and candidate embryo, family's genealogical relationship is grandmother or aunt, and when target gene is on X chromosome, mother effective tagSNP site computing formula is as follows:

$P_{1gt}=\left\{\begin{array}{c}A(P_{gt}=AA)\\ B(P_{gt}=BB)\end{array}\right.---\left(b4\right)$

M _1gt＝R _gt∩M _gt(R _gt≠AB)(c5)

M _2gt＝M _gt-M _1gt(6)

Wherein,

P _1gt: the equipotential type referring to target SNP site in haplotype p1 on autosome or X chromosome, its value is A or B;

P _2gt: the equipotential type referring to target SNP site in haplotype p2 on autosome, its value is A or B, is positioned at the target SNP site on X chromosome, then there is not P _2gt;

M _1gt: the equipotential type referring to target SNP site in haplotype m1 on autosome or X chromosome, its value is A or B;

M _2gt: the equipotential type referring to target SNP site in haplotype m2 on autosome or X chromosome, its value is A or B;

P1: refer to the target gene haplotype on the item chromosome of candidate embryo sample father; P2: refer to the target gene haplotype on the another item chromosome of candidate embryo sample father;

M1: refer to the target gene haplotype on the item chromosome of candidate embryo sample mother; M2: refer to the target gene monomer on the another item chromosome of candidate's Embryos Embryo sample mother;

R _gt: the genotype referring to target SNP site propositus, its value is AA, AB or BB;

P _gt: the genotype referring to target SNP site candidate embryo sample father, its value is AA, AB or BB;

M _gt: the genotype referring to target SNP site candidate embryo sample mother, its value is AA, AB or BB;

A: the base-pair referred in target SNP site is the equipotential type of base A and base T;

B: the base-pair referred in target SNP site is the equipotential type of base C and bases G.

5. the science of heredity pick-up unit before Embryonic limb bud cell as claimed in claim 1, it is characterized in that, in described candidate embryo haplotype reconstruction and genetic origin determination module 300, adopt whether following formula determination target SNP site is crucial SNP site, and obtain each crucial SNP site two chromosomal parental sources on candidate embryo:

1) when family's genealogical relationship of propositus and candidate embryo is elder brother or elder sister, when target gene is on autosome, father effective tagSNP site computing formula is as follows:

E _2O＝“NA”(a7)

$E_{1gt}=\left\{\begin{array}{c}B(M_{gt}=AA,E_{gt}=BB)\\ A(M_{gt}=BB,E_{gt}=AA)\\ E_{gt}-M_{1gt}\end{array}\right.---\left(a8\right)$

$KB=\left\{\begin{array}{c}A(M_{gt}=BB)\\ B(M_{gt}=AA)\end{array}\right.---\left(a10\right)$

Mother effective tagSNP site computing formula is as follows:

E _1O＝“NA”(b9)

$E_{2gt}=\left\{\begin{array}{c}B(P_{gt}=AA,E_{gt}=BB)\\ A(P_{gt}=BB,E_{gt}=AA)\\ E_{gt}-P_{1gt}\end{array}\right.---\left(a12\right)$

$KB=\left\{\begin{array}{c}A(P_{gt}=BB)\\ B(P_{gt}=AA)\end{array}\right.---\left(b10\right)$

2) be man when family's genealogical relationship of propositus and candidate embryo is the sex of elder brother or elder sister and embryo, when target gene is on X chromosome

Mother effective tagSNP site computing formula is as follows:

E _1O＝“NB”(d9)

$E_{2gt}=\left\{\begin{array}{c}A(E_{gt}=AA)\\ B(E_{gt}=BB)\end{array}\right.---\left(b12\right)$

$KB=\left\{\begin{array}{c}A(P_{gt}=BB)\\ B(P_{gt}=AA)\end{array}\right.---\left(b10\right)$

3) sex being elder brother or elder sister and embryo when family's genealogical relationship of propositus and candidate embryo is female, when target gene is on X chromosome

Mother effective tagSNP site computing formula is as follows:

E _1gt＝P _1gt(b8)

E _1O＝“P1”(c9)

$E_{2gt}=\left\{\begin{array}{c}B(P_{gt}=AA,E_{gt}=BB)\\ A(P_{gt}=BB,E_{gt}=AA)\\ E_{gt}-E_{1gt}\end{array}\right.---\left(a12\right)$

$KB=\left\{\begin{array}{c}A(P_{gt}=BB)\\ B(P_{gt}=AA)\end{array}\right.---\left(b10\right)$

4) when family's genealogical relationship of propositus and candidate embryo is grandfather, grandmother, aunt or uncle, when target gene is on autosome

Father effective tagSNP site computing formula is as follows:

E _2O＝“NA”(a7)

$E_{\lg t}=\left\{\begin{array}{c}B(M_{gt}=AA,E_{gt}=BB)\\ A(M_{gt}=BB,E_{gt}=AA)\\ E_{gt}-M_{1gt}\end{array}\right.---\left(a8\right)$

$KB=\left\{\begin{array}{c}A(M_{gt}=BB)\\ B(M_{gt}=AA)\end{array}\right.---\left(a10\right)$

5) when family's genealogical relationship of propositus and candidate embryo is granddad, grandmother, aunt, uncle, when target gene is on autosome

Mother effective tagSNP site computing formula is as follows:

E _1O＝“NA”(b9)

$E_{2gt}=\left\{\begin{array}{c}B(P_{gt}=AA,E_{gt}=BB)\\ A(P_{gt}=BB,E_{gt}=AA)\\ E_{gt}-E_{1gt}\end{array}\right.---\left(a12\right)$

$KB=\left\{\begin{array}{c}A(P_{gt}=BB)\\ B(P_{gt}=AA)\end{array}\right.---\left(b10\right)$

6) when family's genealogical relationship of propositus and candidate embryo be granddad, grandmother, aunt, uncle and embryo sex be man, when target gene is on X chromosome

Mother effective tagSNP site computing formula is as follows:

E _1O＝“NB”(d9)

$E_{2gt}=\left\{\begin{array}{c}A(E_{gt}=AA)\\ B(E_{gt}=BB)\end{array}\right.---\left(b12\right)$

$KB=\left\{\begin{array}{c}A(P_{gt}=BB)\\ B(P_{gt}=AA)\end{array}\right.---\left(b10\right)$

7) when family's genealogical relationship of propositus and candidate embryo be granddad, grandmother, aunt, uncle and embryo sex be female, when target gene is on X chromosome

Mother effective tagSNP site computing formula is as follows:

E _1gt＝P _1gt(b8)

E _1O＝“NA”(b9)

$E_{2gt}=\left\{\begin{array}{c}B(P_{gt}=AA,E_{gt}=BB)\\ A(P_{gt}=BB,E_{gt}=AA)\\ E_{gt}-E_{1gt}\end{array}\right.---\left(a12\right)$

$KB=\left\{\begin{array}{c}A(P_{gt}=BB)\\ B(P_{gt}=AA)\end{array}\right.---\left(b10\right)$

In formula,

E _1O: the parental source of target SNP site on the male parent chromosome referring to candidate embryo, its value is P1, P2, NA or NB;

E _2O: the parental source of target SNP site on the maternal chromosome referring to candidate embryo, its value is M1, M2 or NA;

P1: represent relevant to candidate father embryo, and have genetic association with children propositus or paternal propositus;

P2: represent relevant to candidate father embryo, and with children propositus or paternal propositus without genetic association;

M1: represent relevant to candidate mother embryo, and have genetic association with children propositus or maternal propositus;

M2: represent relevant to candidate mother embryo, and with children propositus or maternal propositus without genetic association;

NA: represent and exist, but cannot to judge or without the need to determining;

NB: represent and do not exist;

E _1gt: the equipotential type referring to target SNP site in haplotype E1 on autosome or X chromosome, its value is A or B; When candidate embryo is the male sex, E _1gtdo not exist;

E _2gt: the equipotential type referring to target SNP site in haplotype E2 on autosome or X chromosome, its value is A or B;

E1: refer to the target gene haplotype of candidate embryo from father; E2: refer to the target gene haplotype of candidate embryo from mother; When candidate embryo is the male sex, E1 does not exist;

E _gt: the genotype referring to candidate embryo, its value is AA, BB or AB, and its value derives from message unit;

KB: refer to crucial equipotential, its value is A or B;

E _r: judge whether target SNP site is crucial SNP site, and value is KeySNP or Non-KeySNP.

6. the science of heredity pick-up unit as claimed in claim 1 before Embryonic limb bud cell, it is characterized in that, described candidate embryo haplotype parental source determination module 400 comprises further:

Candidate embryo haplotype parental source statistical module 410, be connected with candidate embryo haplotype reconstruction and genetic origin determination module 300 and amphiphilic monomer type creation module 200, for the crucial SNP site combination corresponding according to target gene candidate embryo haplotype, and each crucial SNP site two chromosomal parental sources, in statistics target gene candidate embryo haplotype, the crucial SNP site sum that all kinds of crucial SNP site two chromosomal parental sources are corresponding;

Candidate embryo haplotype parental source analysis module 420, be connected with candidate embryo haplotype parental source statistical module 410, for the crucial SNP site sum according to all kinds of parental source of target gene candidate embryo haplotype, determine candidate embryo haplotype parental source.

7. the science of heredity pick-up unit before Embryonic limb bud cell as claimed in claim 6, it is characterized in that, when described candidate embryo haplotype parental source statistical module 410 is also 0 for the crucial SNP site sum corresponding when described all kinds of crucial SNP site two chromosomal parental sources, increase the target SNP site number in the target SNP site combination that in amphiphilic monomer type creation module 200, target gene amphiphilic monomer type is corresponding, until crucial SNP site sum corresponding to described all kinds of crucial SNP site two chromosomal parental sources has at least one not to be 0.

8. the science of heredity pick-up unit before Embryonic limb bud cell as claimed in claim 1, is characterized in that, in described candidate embryo haplotype parental source determination module 400, adopt the parental source of following methods analyst determination target gene candidate embryo haplotype:

1) P _1N>0, P _2N=0, then candidate embryo haplotype E1 parental source is P1;

2) P _1N=0, P _2N>0, then candidate embryo haplotype E1 parental source is P2;

3) P _1N=0, P _2N=0, as each crucial SNP site E _1Owhen being NA, then the parental source of candidate embryo haplotype E1 cannot judge or without the need to determining, be NA; As each crucial SNP site E _1Owhen being NB, then the parental source of candidate embryo haplotype E1 does not exist, and is NB;

4) P _1N>0, P _2N>0, when there is not crucial SNP site in the region at target gene place, the parental source from the nearest crucial SNP site in upstream of target gene and the crucial SNP site in nearest downstream is selected to add up: if the statistics in these two sites is P _1N=2P _2N=0, then candidate embryo haplotype E1 parental source is P1; If statistics is P _2N=2P _1N=0, then candidate embryo haplotype E1 parental source is P2; If statistics is P _1N=1, and P _2N=1, then candidate embryo haplotype E ₁source is P1/P2; When there is crucial SNP site in the region at target gene place, the parental source of the crucial SNP site in the region shared by target gene is added up: if the statistics in these sites is P _1N>0P _2N=0, then candidate embryo haplotype E1 parental source is P1; Statistics is P _1N=0P _2N>0, then candidate embryo haplotype E1 parental source is P2; Statistics is P _1N=0P _2N=0, then the parental source of candidate embryo haplotype E1 is NA or NB; Statistics is P _1N>0P _2N>0, then candidate embryo haplotype E ₁parental source is P1/P2;

5) M _1N>0, M _2N=0, then candidate embryo haplotype E2 parental source is M1;

6) M _1N=0, M _2N=0, then the parental source of candidate embryo haplotype E2 cannot judge, is NA;

7) M _1N=0, M _2N>0, then candidate embryo haplotype E2 parental source is M2;

8) M _1N>0, M _2N>0, when there is not crucial SNP site in the region at target gene place, the parental source from the nearest crucial SNP site in upstream of target gene and the crucial SNP site in nearest downstream is selected to add up: if the statistics in these two sites is M _1N=2M _2N=0, then candidate embryo haplotype E2 parental source is M1; If statistics is M _2N=2M _1N=0, then candidate embryo haplotype E2 parental source is M2; If statistics is M _1N=1, and M _2N=1, then candidate embryo haplotype E2 originates as M1/M2; When there is crucial SNP site in the region at target gene place, the parental source of the crucial SNP site in the region shared by target gene is added up: if the statistics in these sites is M _1N>0M _2N=0, then candidate embryo haplotype E2 parental source is M1, and statistics is M _1N=0M _2N>0, then candidate embryo haplotype E2 parental source is M2, and statistics is M _1N=0M _2N=0, then the parental source of candidate embryo haplotype E2 cannot judge, is NA, and statistics is M _1N>0M _2N>0, then candidate embryo haplotype E2 originates as M1/M2;

In formula:

P _1N: refer in target gene candidate embryo haplotype, the parental source type of crucial SNP site is the crucial SNP site sum of " P1 ";

P _2N: refer in target gene candidate embryo haplotype, the parental source type of crucial SNP site is the crucial SNP site sum of " P2 ";

M _1N: refer in target gene candidate embryo haplotype, the parental source type of crucial SNP site is the crucial SNP site sum of " M1 ";

M _2N: refer in target gene candidate embryo haplotype, the parental source type of crucial SNP site is the crucial SNP site sum of " M2 ".

9. the science of heredity pick-up unit before Embryonic limb bud cell as claimed in claim 1, it is characterized in that, whether whether described candidate embryo carries gene defect judge module 500 also normal for judging the phenotype of candidate embryo.

10. the science of heredity pick-up unit before Embryonic limb bud cell as claimed in claim 9, it is characterized in that, whether candidate embryo carries the pathogenic haplotype in parents and judges that whether the phenotype of candidate embryo is normal to utilize following table relation to judge:

In table,

P1: represent relevant to candidate father embryo, and have genetic association with children propositus or paternal propositus;

P2: represent relevant to candidate father embryo, and with children propositus or paternal propositus without genetic association;

M1: represent relevant to candidate mother embryo, and have genetic association with children propositus or maternal propositus;

M2: represent relevant to candidate mother embryo, and with children propositus or maternal propositus without genetic association;

NA: represent and exist, but cannot to judge or without the need to determining;

NB: represent and do not exist.

Science of heredity pick-up unit before 11. Embryonic limb bud cell as claimed in claim 1, it is characterized in that, the science of heredity pick-up unit before described Embryonic limb bud cell also comprises:

Candidate's embryo chromosome abnormality juding module 600, is connected with message unit 100, for according to the SNP site information of candidate embryo sample and the SNP site information comparison of normal sample, marks the region having chromosome abnormality in candidate embryo.

Science of heredity pick-up unit before 12. Embryonic limb bud cell as claimed in claim 11, is characterized in that having the region of chromosome abnormality specifically to refer to: the B gene frequency band in this region is compared normal phenotype and changed.

Science of heredity pick-up unit before 13. Embryonic limb bud cell as claimed in claim 12, is characterized in that, the B gene frequency band in this region described is compared the situation that normal phenotype changes and is selected from following:

1) in the SNP site B gene frequency figure in this region, the number of band increases, then think that chromosome number increases;

2) in the SNP site B gene frequency figure in this region, the decreased number of band, then think that chromosome number reduces.

Science of heredity pick-up unit before 14. Embryonic limb bud cell as claimed in claim 1, it is characterized in that, described message unit 100 also provides HLA related gene information, and the science of heredity pick-up unit before described Embryonic limb bud cell also comprises:

Candidate embryo HLA distribution type analysis module 700, be connected with message unit 100 and candidate embryo haplotype parental source determination module 400, for with HLA related gene for target gene, according to the parental source of target gene candidate embryo haplotype, filter out the candidate embryo consistent with the parental source of propositus's target gene.

Science of heredity pick-up unit before 15. Embryonic limb bud cell as claimed in claim 14, is characterized in that, the gene that described HLA is relevant comprises HLA-A, HLA-B, HLA-C, HLA-DPB1, HLA-DQB1 and HLA-DRB1.

Science of heredity pick-up unit before 16. Embryonic limb bud cell as claimed in claim 1, it is characterized in that, science of heredity pick-up unit before described Embryonic limb bud cell also comprises: sample Quality Control unit 800, be connected with message unit 100, for judging whether the recall rate of the SNP of candidate embryo sample father, the SNP site of candidate embryo sample mother, the SNP site of propositus, the SNP site of candidate embryo exceedes threshold value, and judge whether the sex input of candidate embryo sample parents' sex and propositus is wrong.