KR101774996B1 - Markers For Predicting Effect of Therapeutic Agent Against Hemophilia and Use Thereof - Google Patents

Abstract

The present invention relates to a marker for predicting efficacy of a hemophilia treatment agent. More specifically, the present invention relates to a single nucleotide polymorphism (SNP) marker for predicting efficacy of a hemophilia treatment agent, a composition containing a preparation capable of detecting the SNP marker, a microarray or a kit containing the composition, a method for providing information for predicting efficacy of the hemophilia treatment agent using the SNP marker, and a method for selecting SNP for predicting efficacy of the hemophilia treatment agent.

Description

Markers for Predicting Effect of Therapeutic Agents Against Hemophilia and Use Thereof

The present invention relates to a marker for predicting the efficacy of a therapeutic agent for hemophilia, and more particularly to a composition comprising a single nucleotide polymorphism (SNP) marker for predicting the efficacy of a therapeutic agent for hemophilia, an agent capable of detecting the SNP marker, Or a microarray, a method for providing information for predicting the efficacy of a hemophilia therapeutic agent using the SNP marker, and a method for selecting a SNP for predicting efficacy of a hemophilia therapeutic agent.

Hemophilia is a haemorrhagic disease caused by a deficiency of blood clotting factors. Factor VIII deficiency is referred to as hemophilia A, and Factor IX deficiency as hemophilia B. The incidence of haemophilia A occurs from 5,000 to 1 in 10,000, and the incidence of haemophilia B is estimated to be about one-fifth of that (Park YS, 2009; 52 (12): 1201- 6). Hemophilia is classified as severe (<1%), moderate (1-5%) or mild (> 5%) depending on the activity of blood coagulation factors. In general, hemophilia therapy replenishes deficient blood coagulation factors. Currently Factor VIII, Factor IX agents are used, and if antibodies are produced during treatment, bypass agents are administered (Kemton CL et al., Blood. 2009; 113 ): 11-7).

It is very important to predict the inhibitor development response in hemophilia drug therapy. The incidence of antibody production in hemophilia A patients is about 30%, and about 3% in hemophilia B patients (Kessler CM, Hematology Am Soc Hematol Educ. Program 2005: 429-35). Antibodies to hemophilia therapy are most commonly encountered within 50 days of drug exposure, and are classified as antibody-producing when they are more than 0.6 Bethesda Unit (BU), and are classified as high antibody-producing when they are above 5 BU (Kasper CK et al. , Thromb Diath Haemorrh 1975; 34 (2): 612; Verbruggen B et al., Thromb Haemost 1995: 73 (2): 247-51; Viel KR et al., N Engl J Med. ): 1618-27; Kemton CL et al., Blood. 2009; 113 (1): 11-7). However, there is a high incidence of joint complications due to frequent bleeding (Park YS, J Korean Med Assoc 2009; 52 (12): 1201-6). Alternative therapies include bypass agents such as activated prothrombin complexes and Factor VII recombinant agents, but it may be more desirable to remove the antibody through immunotherapy in the antibody-generated response group (Kemton CL et al., Blood. 2009 ; 113 (1): 11-7; Park YS, J Korean Med Assoc 2009; 52 (12): 1201-6). The success rate of immune tolerance varies from 30 to 80%, and patients can continue to use Factor VIII or Factor IX after immunotherapy (Park YS, J Korean Med Assoc 2009; 52 (12): 1201- 6).

A number of studies have been conducted on the major predictors or causes of antibody production in hemophilia therapy. The genetic predisposition factor is known to be the most likely cause of defects in the F8 gene (Schwaab R et al., 1995 [6]: 1402-6; Oldenburg J et al., Thromb Haemost 1997, SNP (single nucleotide polymorphism) mutations located in genes such as MHC II, TNF-α, IL-10 and CTLA-4 are also statistically significant 1997). In addition, a number of genetic predictors are expected to be present (Hay CR et al., 1997 (2): 234-7; Oldenburg J et al., Thromb Haemost. Astmar J et al., J Thromb Haemost. 2007 (2006), 379-45; Astark J et al., 2006, 107 (8): 3167-72; , 5 (2): 263-5).

Therefore, the inventors of the present application conducted a genome wide association study using Exome Sequencing data to identify the genetic prediction marker of the antibody production reaction of hemophilia therapy, , And finally completed the present invention.

It is an object of the present invention to provide an efficacy predictive SNP marker of hemophilia therapy.

Another object of the present invention is to provide a composition for predicting the efficacy of a therapeutic agent for hemophilia comprising an agent capable of detecting the predictive SNP marker of the hemophilia therapeutic agent.

Another object of the present invention is to provide a kit or a microarray for predicting the efficacy of a therapeutic agent for hemophilia comprising the composition for predicting efficacy of the therapeutic agent for hemophilia.

Yet another object of the present invention is to provide a method for providing information for predicting the efficacy of a therapeutic agent for hemophilia comprising the step of determining a base at a polymorphic site of the SNP marker.

Another object of the present invention is to provide a method for screening SNP markers for predicting the efficacy of a therapeutic agent for hemophilia.

In order to accomplish the above object, the present invention provides a nucleic acid molecule comprising a nucleotide sequence derived from the nucleotide sequence of SEQ ID NOS: 1 to 87 and comprising at least 5 consecutive nucleotides comprising the nucleotide sequence 101 of SEQ ID NO: 1 to 84 and the nucleotide sequence of SEQ ID NO: 85 to 87, A single nucleotide polymorphism (SNP) marker for the hemophilia therapeutic comprising one or more polynucleotides selected from polynucleotides composed of a nucleotide or a complementary polynucleotide thereof.

The present invention also provides a composition for predicting efficacy of a therapeutic agent for hemophilia comprising a probe, a primer or an extramamer capable of detecting the SNP marker.

The present invention also provides an efficacy prediction kit or microarray for hemophilia therapy comprising the composition.

(A) amplifying or hybridizing a polymorphic site of the SNP marker with a probe from the DNA of the isolated sample; And (b) determining a base of the amplified or hybridized polymorphic site of step (a). The present invention also provides a method for providing information for predicting the efficacy of a therapeutic agent for hemophilia.

The present invention also relates to a method for screening for SNPs in which (a) SNPs having a sequencing depth value of 10 or more and a genotype quality value of 20 or more are selected in wholeexome sequencing of a subject to which a hemophilia treatment agent is administered , Removing the intergenic variant in which there is no residual region in the periphery; (b) performing a linkage analysis on the selected SNPs in step (a), thereby selecting SNPs that are related to the antibody-producing phenotype of hemophilia treatment; And (c) selecting a SNP in a gene region related to the immune system for the SNP selected in the step (b).

According to the present invention, an objective risk of antibody production to a hemophilia therapeutic agent can be predicted by using the predictive Efficacy SNP marker of hemophilia treatment agent.

FIG. 1 is a schematic view showing a method of selecting an effect predictive SNP marker of a hemophilia therapeutic agent.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein and the experimental methods described below are well known and commonly used in the art.

The present invention, in one aspect, provides a single nucleotide polymorphism (SNP) marker that is a predictive single nucleotide polymorphism for the efficacy of hemophilia therapy.

The marker is preferably selected from the group consisting of 5 or more consecutive bases derived from the nucleotide sequences of SEQ ID NOS: 1 to 87 and comprising the 101st base of SEQ ID NOS: 1 to 84 and the 100th base of SEQ ID NOS: 85 to 87 as SNPs A single nucleotide polymorphism (SNP) marker of the hemophilia therapeutic comprising one or more polynucleotides selected from polynucleotides or their complementary polynucleotides.

In the present invention, "hemophilia" is a haemorrhagic disease caused by a deficiency of blood coagulation factor, and Factor VIII deficiency is termed hemophilia A, and Factor IX deficiency is termed hemophilia B. Type A haemophilia is a hemorrhagic disease characterized by a quantitative or qualitative defect of FVIII due to defect of FVIII gene and type B haemophilia is characterized by a quantitative or qualitative defect of FIX due to abnormality of FIX gene.

Preferably, the present invention relates to the prediction of the efficacy of the hemophilia A therapeutic agent.

The term "polymorphism " used in the present invention refers to the case where two or more alleles exist in one locus, and among the polymorphic sites, It is called single nucleotide polymorphism (SNP). Preferred polymorphic markers have two or more alleles with a frequency of occurrence of 1% or more, more preferably 5% or 10% or more in the selected population.

As used herein, the term "allele" refers to various types of a gene present at the same gene locus on a homologous chromosome. Alleles are also used to represent polymorphisms, for example, SNPs have two kinds of bialles.

The term " rs_id " used in the present invention means an independent marker rs-ID assigned to all SNPs initially registered by the NCBI that has started to accumulate SNP information since 1998. [ In the present invention, it is described in the form of rs2486316. The rs_id shown in the above table means the SNP marker which is the polymorphism marker of the present invention. Those skilled in the art will readily be able to ascertain the location and sequence of the SNP using the rs_id. The specific sequence corresponding to rs_id, the number of the NCBI's Single Nucleotide Polymorphism Database (dbSNP), may change slightly over time. It will be apparent to those skilled in the art that the scope of the present invention also affects the altered sequence.

The above "SEQ ID NOS: 1 to 87" is a polymorphic sequence including a polymorphic site. A polymorphic sequence means a sequence comprising a polymorphic site comprising a SNP in a polynucleotide sequence. The polynucleotide sequence may be DNA or RNA.

The SNP markers of the present invention are the markers shown in Tables 1 and 2, and when the polymorphic site of the SNP marker of an individual is an allele indicated by a risk allele, it is determined that the individual is at an increased risk of antibody production to the hemophilia treatment And alleles marked with a protective allele can be considered as individuals with a low risk of antibody production to the hemophilia treatment. In the present invention, "risk allele" may be used in combination with "effect allele ".

Preferably, the SNP marker is an EFFECT predictive SNP marker of a hemophilia therapeutic agent. In the polynucleotide derived from the nucleotide sequence of SEQ ID NO: 1 to 87, the SNP marker of SEQ ID NO: 1 to 84 Base or a polynucleotide whose 100 th base of SEQ ID NOS: 85 to 87 is composed of 5 or more consecutive bases comprising allelic bases of Tables 1 and 2, or a complementary polynucleotide thereof, Lt; / RTI &gt;

Efficacy prediction of hemophilia therapy according to the present invention The SNP may be composed of each of the SNPs, more preferably 2 or more SNPs, and most preferably all of the SNPs 1 to 87.

It is preferable that the polynucleotide or the complementary polynucleotide thereof for predicting the efficacy of a therapeutic agent for hemophilia comprising the SNP according to the present invention is composed of 5 or more continuous bases, more preferably 5 to 50 More preferably from 10 to 40, even more preferably from 20 to 30 contiguous bases, but is not limited thereto.

In another aspect, the present invention provides a composition for predicting efficacy of a therapeutic agent for hemophilia comprising an agent capable of detecting the SNP marker.

In the present invention, the detectable agent may be a probe. The probe refers to an oligonucleotide which can be identified through a hybridization reaction specifically with a polymorphic site of the above-mentioned gene to predict the efficacy of the therapeutic agent for hemophilia.

The allele-specific probe of the present invention has a polymorphic site among nucleic acid fragments derived from two individuals of the same species, and hybridizes to a DNA fragment derived from one individual, but not to a fragment derived from another individual. In this case, the hybridization conditions show a significant difference in the hybridization intensity between the alleles, and should be sufficiently strict so that only one of the alleles hybridizes. Preferably, the probe of the present invention aligns with the polymorphic site of the polymorphic sequence. This can lead to good hybridization differences between different allelic forms. The probe of the present invention can be used for a diagnostic kit or a diagnostic method such as a microarray for detecting alleles and diagnosing sensitivity to the efficacy of a hemophilia therapeutic. The specific method of such gene analysis is not particularly limited, and may be by any gene detection method known in the art.

In the present invention, the detectable agent may be a primer. The primer of the present invention refers to a composition capable of identifying the polymorphic site of the above-mentioned gene through amplification to predict the efficacy of the therapeutic agent for hemophilia. Preferably, the primer specifically amplifies the polynucleotide of the efficacy prediction marker of the hemophilia therapeutic agent Primer &quot;

The primers used for the polymorphic marker amplification can be amplified by PCR using appropriate conditions in suitable buffers (e.g., 4 different nucleoside triphosphates and polymerases such as DNA polymerase) and at a suitable temperature, which can serve as a starting point for template- Refers to single stranded oligonucleotides. The appropriate length of the primer may vary depending on the intended use, but is usually 15 to 30 nucleotides. Short primer molecules generally require a lower temperature to form a stable hybrid with the template. The primer sequence need not be completely complementary to the template, but should be sufficiently complementary to hybridize with the template. The primer hybridizes to a DNA sequence containing a polymorphic site to amplify a DNA fragment containing the polymorphic site.

As used herein, the term "primer" refers to a base sequence having a short free 3 'hydroxyl group, capable of forming a base pair with a complementary template, Quot; means a short sequence functioning as a starting point for &lt; / RTI &gt; The primers can initiate DNA synthesis in the presence of reagents and four different nucleoside triphosphates for polymerization reactions (i. E., DNA polymerase or reverse transcriptase) at appropriate buffer solutions and temperatures. PCR amplification can be used to predict whether antibodies to the hemophilia treatment will be produced through the production of desired products. The PCR conditions, the lengths of the sense and antisense primers can be modified based on what is known in the art.

The probes or primers of the present invention can be chemically synthesized using the phosphoramidite solid support method, or other well-known methods. Such nucleic acid sequences may also be modified using many means known in the art. Non-limiting examples of such modifications include, but are not limited to, methylation, "capping ", replacement of natural nucleotides with one or more homologues, and modifications between nucleotides, such as uncharged linkers, such as methylphosphonate, Phosphoamidates, carbamates, etc.) or charged linkages (e.g., phosphorothioates, phosphorodithioates, etc.).

In the present invention, the above-mentioned detectable agent may be an aptamer.

Aptamer is a single chain DNA or RNA molecule, which refers to a small single stranded oligonucleotide capable of specifically recognizing a target substance with high affinity. Aptamers can be used as a component of a biosensor capable of recognizing molecules in a detection assay system and have been recognized as a substitute for antibodies. In particular, aptamers can be used as target molecules for various organic and inorganic substances, including toxins, unlike antibodies, and once an extramammer that specifically binds to a specific substance is isolated, automated oligomer synthesis methods can be used to reproduce This was economically feasible. Since the development of an abutamer-based biosensor measuring the target protein using the fluorescent-labeled abstammer for the first time in 1996, various types of abstamator biosensors have been developed based on the advantages and structural characteristics of the abutmenter (Kim, Yeon-seok & Guman-bok, NICE, 26 (6): 690, 2008).

In another aspect, the present invention provides a kit for predicting the efficacy of a therapeutic agent for hemophilia comprising the composition for predicting efficacy of the therapeutic agent for hemophilia.

The kit may be a PCR kit, a DNA analysis kit (for example, a DNA chip), or a fluorescence dye kit, but is not limited thereto.

The kit of the present invention can predict the efficacy of the hemophilia treatment agent by confirming the SNP marker, which is an effect prediction marker of the hemophilia therapeutic agent, by amplification or by confirming the SNP marker in the amplified cDNA template for mRNA.

As a specific example, the kit of the present invention comprises a genomic DNA derived from a sample to be analyzed, a primer set specific for the marker of the present invention, an appropriate amount of DNA polymerase (for example, Taq polymerase), dNTP Mixture, PCR buffer solution and water. The PCR buffer solution may contain KCl, Tris-HCl and MgCl2. In addition, components necessary for conducting electrophoresis to confirm whether the PCR product is amplified can be further included in the kit of the present invention.

In addition, the kit of the present invention may be an efficacy prediction kit of a hemophilia therapy agent including elements necessary for performing a fluorescent dye kit using a probe type. As a method of analysis using a dyestuff dye, TaqMan 5 '(nuclease assay), molecular beacons, OLA (oligonucleotide ligase assay), Invader Assay (Invasive Cleavage of Oligonucleotide Probes), Pyrosequencing, Single Base Extension or Fluorescence Polarization , But is not limited thereto.

Also preferably, the kit of the present invention may be an efficacy prediction kit of a hemophilia therapeutic agent containing essential elements necessary for performing a DNA chip. DNA chip kits are those in which nucleic acid species are attached in a gridded array on a generally flat solid support plate, typically a glass surface not larger than a slide for a microscope, and nucleic acids are uniformly arranged on the chip surface, Hybridization reaction occurs between the nucleic acid on the surface and the complementary nucleic acid contained in the solution treated on the surface of the chip to enable a mass parallel analysis.

In another aspect, the present invention provides an efficacy prediction microarray of a hemophilia therapeutic agent comprising a polynucleotide of an efficacy prediction marker of the hemophilia therapeutic agent.

The microarray may comprise DNA or RNA polynucleotides. The microarray may be a conventional microarray, except for including the polynucleotides, polypeptides, cDNA, etc. of the present invention.

Preferably, the hemophilia therapy antagonist is a polynucleotide capable of hybridizing specifically to a base of a polymorphic site in each polymorphic sequence of SEQ ID NOS: 1 to 87, a complementary polynucleotide thereof, , A polynucleotide that hybridizes with the polynucleotide of the present invention, a polypeptide encoded by the polynucleotide, or cDNA thereof, as a probe, can be produced by a conventional method known to those skilled in the art.

The polynucleotide is preferably immobilized on a substrate coated with one activator selected from the group consisting of amino-silane, poly-L-lysine and aldehyde But is not limited thereto.

Methods for producing microarrays by immobilizing probe polynucleotides on a substrate are well known in the art. The probe polynucleotide means a polynucleotide capable of hybridizing, and means an oligonucleotide capable of binding to the complementary strand of the nucleic acid in a sequence-specific manner. The probe of the present invention is an allele-specific probe in which a polymorphic site exists in a nucleic acid fragment derived from two members of the same species and hybridizes to a DNA fragment derived from one member but does not hybridize to a fragment derived from another member . In this case, the hybridization conditions show a significant difference in the intensity of hybridization between alleles, and should be sufficiently stringent to hybridize to only one of the alleles. This can lead to good hybridization differences between different allelic forms. The probe of the present invention can be used for predicting the efficacy of a therapeutic agent for hemophilia by detecting an allele. The prediction method includes detection methods based on hybridization of a nucleic acid such as a Southern blot, and may be provided in a form pre-bonded to a substrate of a DNA chip in a method using a DNA chip.

The process of immobilizing the probe polynucleotide on the substrate associated with the prediction of the efficacy of the hemophilia therapeutic of the present invention can also be easily produced using such a conventional technique. The method of immobilizing the polynucleotide on the substrate is preferably a micropipetting method using a piezo electric method or a method using a pin type spotter, It does not.

In addition, hybridization of nucleic acids on a microarray and detection of hybridization results are well known in the art. The detection can be accomplished, for example, by labeling the nucleic acid sample with a labeling substance capable of generating a detectable signal comprising a fluorescent substance, such as Cy3 and Cy5, and then hybridizing on the microarray and detecting The hybridization result can be detected by detecting the generated signal.

In another aspect, the present invention provides a method for providing information for predicting the efficacy of a hemophilic therapeutic agent, comprising determining a base at a polymorphic site of the SNP marker.

According to an embodiment of the present invention, there is provided a method for amplifying a polynucleotide comprising: (a) amplifying a polymorphic site of the SNP marker from DNA of a separated sample or hybridizing with a probe; And (b) determining the base of the amplified or hybridized polymorphic site of step (a).

The DNA of the isolated sample can be obtained from a sample isolated from the individual.

The term "individual" of the present invention refers to a subject who predicts the efficacy of a therapeutic agent for hemophilia. The prediction of the efficacy of the therapeutic agent for hemophilia is based on the odds ratio or relative risk of antibody production in the hemophilia therapy- . DNA can be obtained from the sample, such as hair, urine, blood, various body fluids, isolated tissues, isolated cells or saliva, but is not limited thereto.

The step of amplifying the polymorphic site of the SNP marker from the DNA of step (a) or hybridizing with the probe may be performed by any method known to those skilled in the art. For example, the target nucleic acid can be obtained by PCR amplification and purification thereof. Other ligase chain reaction (LCR), transcription amplification and self-sustaining sequence replication and nucleic acid based sequence amplification (NASBA) can be used.

Determination of bases in the polymorphic site of step (b) of the above method can be carried out by sequencing, hybridization by microarray, allele specific PCR, dynamic allele-specific hybridization , DASH), PCR extension analysis, PCR-SSCP, PCR-RFLP analysis or TaqMan technique, SNPlex platform (Applied Biosystems), mass spectrometry (eg, Sequenom's MassARRAY system), mini- But are not limited to, the Plex system (BioRad), the CEQ and SNPstream system (Beckman), the Molecular Inversion probe array technology (e.g. Affymetrix GeneChip), and BeadArray Technologies (e.g. Illumina GoldenGate and Infinium analysis) . One or more alleles in a polymorphic marker can be identified, including microsatellite, SNP, or other types of polymorphic markers, by such methods or by other methods available to those skilled in the art to which this invention belongs. The base of such a polymorphic site can be determined preferably through a SNP chip.

The term "SNP chip" used in the present invention means one of DNA microarrays capable of confirming each base of several hundred thousand SNPs at a time.

The TaqMan method comprises the steps of: (1) designing and constructing a primer and a TaqMan probe to amplify a desired DNA fragment; (2) labeling probes of different alleles with FAM dyes and VIC dyes (Applied Biosystems); (3) performing PCR using the DNA as a template and using the primer and the probe; (4) after completion of the PCR reaction, analyzing and confirming the TaqMan assay plate with a nucleic acid analyzer; And (5) determining the genotype of the polynucleotides of step (1) from the analysis result.

In the above, the sequencing analysis can be performed using a conventional method for determining the nucleotide sequence, and can be performed using an automated gene analyzer. The allele-specific PCR means a PCR method in which a DNA fragment in which the SNP is located is amplified with a primer set including a primer designed with the base at the 3 'end at which the SNP is located.

PCR extension analysis is performed by first amplifying a DNA fragment containing a base in which a single base polymorphism is located with a pair of primers and then inactivating all the nucleotides added to the reaction by dephosphorylation and adding SNP specific extension primer, dNTP mixture , Digoxinucleotide, reaction buffer, and DNA polymerase to perform primer extension reaction. At this time, the extension primer has a base immediately adjacent to the 5 'direction of the base in which the SNP is located at the 3' terminus, and the nucleic acid having the same base as the dodecoxynucleotide is excluded in the dNTP mixture, and the dodecoxynucleotide indicates the SNP Base type. As the detection method, when the extension primer or the dioxynucleotide is fluorescently labeled, the SNP is detected by detecting fluorescence using a gene analyzer (for example, Model 3700 of ABI Co., Ltd.) used for general nucleotide sequence determination And when the unlabeled extension primer and the didyxin nucleotide are used, the SNP can be detected by measuring the molecular weight using MALDI-TOF (matrix assisted laser desorption ionization-time of flight) technique.

Among the nucleotide sequences determined in the step (b), when the 101st base is T in rs2486316 described in SEQ ID NO: 1; In rs476488 described in SEQ ID NO: 2, when the 101st base is C; In rs55700376 described in SEQ ID NO: 3, when the 101st base is A; In rs1131568 described in SEQ ID NO: 4, when the 101st base is T; In rs3815496 described in SEQ ID NO: 5, when the 101st base is C; In rs62334652 described in SEQ ID NO: 6, when the 101st base is C; In rs45468202 described in SEQ ID NO: 7, when the 101st base is C; In rs3805220 described in SEQ ID NO: 8, when the 101st base is A; In rs13306436 described in SEQ ID NO: 9, when the 101st base is A; In rs2307075 described in SEQ ID NO: 10, when the 101st base is C; In the case of rs703 represented by SEQ ID NO: 11, when the 101st base is C; In rs16928973 described in SEQ ID NO: 12, when the 101st base is T; In rs2074226 described in SEQ ID NO: 13, when the 101st base is T; In rs10844140 described in SEQ ID NO: 14, when the 101st base is T; In rs2074572 described in SEQ ID NO: 15, when the 101st base is T; In rs9367 described in SEQ ID NO: 16, when the 101st base is C; In the case of rs3816553 described in SEQ ID NO: 17, when the 101st base is A; In rs383483 described in SEQ ID NO: 18, when the 101st base is G; In rs2227982 described in SEQ ID NO: 19, when the 101st base is A; In rs73019023 described in SEQ ID NO: 20, when the 101st base is G; In rs75345202 described in SEQ ID NO: 21, when the 101st base is A; In rs2287872 described in SEQ ID NO: 22, when the 101st base is A; In the case of rs3815734 described in SEQ ID NO: 23, when the 101st base is C; In rs2239354 represented by SEQ ID NO: 24, when the 101st base is A; In rs8099222 described in SEQ ID NO: 25, when the 101st base is A; In the case of rs6136376 described in SEQ ID NO: 26, when the 101st base is A; In rs190983971 described in SEQ ID NO: 58, when the 101st base is G; In rs117064599 described in SEQ ID NO: 59, when the 101st base is G; In rs12091406 described in SEQ ID NO: 60, when the 101st base is A; In rs117447527 described in SEQ ID NO: 61, when the 101st base is A; In rs3738869 represented by SEQ ID NO: 62, when the 101st base is C; In rs200662898 described in SEQ ID NO: 63, when the 101st base is T; In rs17878995 described in SEQ ID NO: 64, when the 101st base is T; In rs140930246 described in SEQ ID NO: 65, when the 101st base is C; In rs75953973 described in SEQ ID NO: 66, when the 101st base is A; In rs149045171 described in SEQ ID NO: 67, when the 101st base is A; In rs732072 described in SEQ ID NO: 68, when the 101st base is A; In rs72481820 described in SEQ ID NO: 69, when the 101st base is T; In rs78245838 described in SEQ ID NO: 70, when the 101st base is C; In rs117884186 described in SEQ ID NO: 71, when the 101st base is C; In the nucleotide sequence shown in SEQ ID NO: 85, when the 100th base is G; In the nucleotide sequence shown in SEQ ID NO: 86, when the 100th base is A; Or in the nucleotide sequence shown in SEQ ID NO: 87, when the 100th base is C, it can be judged that the risk of antibody production to the hemophilia therapeutic agent increases. The polymorphic site is judged to be a risk allele, and the higher the frequency of the allele gene, the higher the risk of the antibody production of the hemophilia therapeutic agent can be determined. More preferably, if two or more polymorphic sites of SEQ ID NOS: 1 to 26, 58 to 71, and 85 to 87 are risk alleles, and most preferably all 43 polymorphisms are risk alleles, . If the 101 &lt; th &gt; base is not the base of the risk allele in Table 1, it can be judged that the risk of the antibody production of the hemophilia therapeutic is low.

The SNPs in Table 1 (OR> 1) increase the risk of antibody production to hemophilia treatment.

More preferably, if one or more, more preferably two or more of the polymorphic sites of SEQ ID NOS: 1, 9, 58 to 71, 85 and 86 are risk alleles, most preferably all 18 are risk alleles It can be judged that there is a high risk of antibody production of hemophilia A therapeutic agent. When the 101st base of SEQ ID NOS: 1, 9, 58 to 71 and the 100th base of SEQ ID NOS: 84 and 85 are not bases of the effect allele of Table 1, it can be judged that the risk of the antibody production of the hemophilia therapeutic is low.

In addition, in the case of rs366316 of SEQ ID NO: 27 among the nucleotide sequences determined in the step (b), when the 101st base is G; In rs138067550 described in SEQ ID NO: 28, when the 101st base is T; In the case of rs3213479 described in SEQ ID NO: 29, when the 101st base is A; In the case of rs17141154 described in SEQ ID NO: 30, when the 101st base is A; In rs2269443 described in SEQ ID NO: 31, when the 101st base is A; In rs2459467 described in SEQ ID NO: 32, when the 101st base is T; In rs2230525 described in SEQ ID NO: 33, when the 101st base is C; In rs6876997 described in SEQ ID NO: 34, when the 101st base is G; In rs149956505 described in SEQ ID NO: 35, when the 101st base is T; In rs2504020 described in SEQ ID NO: 36, when the 101st base is G; In rs2252752 described in SEQ ID NO: 37, when the 101st base is C; In rs2298584 described in SEQ ID NO: 38, when the 101st base is G; In rs4762088 described in SEQ ID NO: 39, when the 101st base is C; In rs3024610 described in SEQ ID NO: 40, when the 101st base is T; In rs1985 described in SEQ ID NO: 41, when the 101st base is A; In rs3765622 described in SEQ ID NO: 42, when the 101st base is G; In rs3826637 described in SEQ ID NO: 43, when the 101st base is A; In rs7226624 described in SEQ ID NO: 44, when the 101st base is C; In rs2240592 described in SEQ ID NO: 45, when the 101st base is C; In rs2252673 described in SEQ ID NO: 46, when the 101st base is G; In rs55882956 described in SEQ ID NO: 47, when the 101st base is A; In rs56374127 described in SEQ ID NO: 48, when the 101st base is T; In rs229525 described in SEQ ID NO: 49, when the 101st base is G; In rs144426992 described in SEQ ID NO: 50, when the 101st base is G; In rs1465922 described in SEQ ID NO: 51, when the 101st base is A; In the case of rs2112290 represented by SEQ ID NO: 52, when the 101st base is A; In rs2072372 described in SEQ ID NO: 53, when the 101st base is A; In rs10844141 described in SEQ ID NO: 54, when the 101st base is G; In rs71359461 described in SEQ ID NO: 55, when the 101st base is C; In rs2305372 described in SEQ ID NO: 56, when the 101st base is T; In rs1057114 described in SEQ ID NO: 57, when the 101st base is C; In rs139568272 described in SEQ ID NO: 72, when the 101st base is A; In rs2298022 described in SEQ ID NO: 73, when the 101st base is A; In rs13097123 described in SEQ ID NO: 74, when the 101st base is T; In rs35625434 represented by SEQ ID NO: 75, when the 101st base is A; In rs185470785 described in SEQ ID NO: 76, when the 101st base is G; In rs61332891 described in SEQ ID NO: 77, when the 101st base is T; In the case of rs3793744 represented by SEQ ID NO: 78, when the 101st base is A; In rs17154981 described in SEQ ID NO: 79, when the 101st base is A; In rs77893724 described in SEQ ID NO: 80, when the 101st base is C; In rs55851358 described in SEQ ID NO: 81, when the 101st base is C; In rs11545564 described in SEQ ID NO: 82, when the 101st base is G; In rs139142355 described in SEQ ID NO: 83, when the 101st base is T; Or rs56292769 described in SEQ ID NO: 84, when the 101st base is C, it can be judged that the risk of antibody production to the hemophilia treatment is decreased. The polymorphic site is judged to be a protective allele, and the higher the frequency of the allele gene, the lower the risk of antibody production in the hemophilia therapy. More preferably, when two or more polymorphic sites of SEQ ID NOS: 27 to 57 and 72 to 84 are protective alleles, most preferably all of the 44 polymorphic sites are considered to have a lower risk of producing hemophilia A therapeutic antibody. If the 101 &lt; th &gt; base is not the base of the protective allele of Table 2, it can be judged that the risk of the antibody production of the hemophilia therapeutic is high.

SNP alleles with OR <1 observed statistically significant Genotype Genotype SEQ ID NO: Reference Opposition SEQ ID NO: Reference Opposition 27 A G 49 A G 28 C T 50 A G 29 G A 51 G A 30 G A 52 G A 31 G A 53 C A 32 A T 54 A G 33 T C 55 G C 34 A G 56 A T 35 A T 57 G C 36 C G 72 G A 37 T C 73 G A 38 A G 74 C T 39 G C 75 G A 40 C T 76 A G 41 T A 77 A T 42 C G 78 G A 43 G A 79 G A 44 T C 80 A C 45 G C 81 T C 46 C G 82 A G 47 G A 83 G T 48 C T 84 G C

The SNP (OR < 1) in Table 2 reduces the risk of antibody production to hemophilia treatment.

More preferably, at least one polymorphic site in SEQ ID NOS: 72 to 84, more preferably two or more polymorphic sites, is most likely to be a risk allele, and most preferably all 13 polymorphic sites have a lower risk of antibody production can do. If the 101 &lt; th &gt; base is not the base of the protective allele of Table 2, it can be judged that the risk of the antibody production of the hemophilia therapeutic is high.

In another aspect, the present invention provides a method for screening the efficacy-predictive SNP of a hemophilia therapeutic.

According to one embodiment of the present invention, there is provided a method of detecting hemophilia comprising the steps of (a) subjecting a subject to hemophilia treatment with a sequencing depth value of 10 or more and a genotype quality value of 20 or more in whole exome sequencing Selecting an SNP, and removing an intergenic variant in which no residue region exists; (b) performing a linkage analysis on the selected SNPs in step (a), thereby selecting SNPs that are related to the antibody-producing phenotype of hemophilia treatment; And (c) selecting a SNP in a gene region related to the immune system for the SNP selected in the step (b).

The term "Sequencing Depth" refers to the number of bases read during the sequencing process. The sequencing depth (range) indicates how many times the total number read out is several times longer than the length of the sequence being analyzed, and the depth is the average number of reads that represent a particular base in the reconstructed sequence. In addition, the term GQ (Genotype Quality) is a Phred-scaled value indicating the reliability that the genotype is a genotype. The higher the value, the more accurate the genotype.

Whole exome sequencing of step (a) above can be performed on a NextSeq 500 instrument using a SureSelect XT v5 Kit for DNA samples of subjects undergoing hemophilia treatment. "Intergenic strain" means a SNP in which no gene region exists in the vicinity of the selected SNPs by performing the full-length exocytic base sequence analysis.

In the step (b), the correlation analysis is performed using Fisher's Exact Test, which is a method of testing the frequency data constituting the partition table using an initial probability distribution. According to one embodiment of the present invention, association analysis is performed for the whole population, F8 inversion detection group, and the group comparison for antibody generation reaction is> 0.60 BU group vs. 0.60 BU group and> 5.00 BU group vs. 5.00 BU group. Although the step (b) is not limited thereto, it is possible to first select SNPs satisfying P &lt; 0.05 with an OR value of &gt; 5 or <0.2 with respect to the whole group (n = 250) SNPs in patients with hemophilia A can be screened only for SNPs with repeated statistical significance for the whole population or the F8 inversion detection group.

The term "OR value" used in the present invention refers to the odds ratio, which is a ratio calculated as an estimate of relative risk in a case-control study. In the case of the control group, Represents the ratio of the probability of having the corresponding SNP in the group. For example, if a person exposed to a risk factor, a person who has not exposed a person, a person who has exposed a person in a control group, or a person who is a non-exposed person in the case group, the ozone ratio is displayed as ad / bc.

If the odds ratio is greater than 1, the corresponding multiple SNPs are associated with antibody production to hemophilia therapy. If the odds ratio is less than 1, the multiple SNPs are not associated with antibody production to the hemophilia A treatment. The larger the odds ratio, the greater the association, and the smaller the odds ratio is, the lower the relevance.

The results of the SNP polymorphism can also be statistically processed using statistical analysis methods commonly used in the art. For example, Student's t-test, Chi- continuous variables, categorical variables, and 95% confidence intervals (CIs) obtained through linear regression analysis, linear regression line analysis, and multiple logistic regression analysis. ) Can be used for analysis.

In the step (c), the SNPs of the gene region related to the immune system are selected for the SNP selected in the step (b), and preferably the HGVS (Human Genome Variation Society) information of the corresponding SNP is confirmed. According to an embodiment of the present invention, 87 SNP marker sets (4 redundant SNPs) for predicting the efficacy of hemophilia therapy were selected by performing steps (a) through (c).

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these examples are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.

Example 1: Tests for production samples and antibodies against hemophilia

(1) Subject selection

A total of 250 hemophilic patients were treated with the SureSelect XT v5 Kit for DNA samples (F8 inversion detection: 141, F8 inversion not detected: 109), and Whole Exome Sequencing was performed on NextSeq 500 Respectively.

(2) Hemophilia treatment antibody production test

In the hemophilia drug treatment, the number of patients with F8 antibody titer was 0.60 BU or less, 108 patients, and 0.61 BU to 5.00 BU and 78 patients, respectively. The composition of the working sample group can be shown in Table 3.

The composition of the working sample ≤ 0.60 BU (group 1) 0.61 to 5.00 BU (group 2) ≥ 5.01 BU (group 3) all
(n = 250) 108 people 78 people 64 people F8 inversion detection
(n = 141) 63 people 54 people 24 people

Example 2: Construction of a SNP set

(1) Whole Exome Sequencing

In order to establish the predictive mutation set of antibody production reaction hemophilia therapeutic agent, 3 steps were performed in total. In this step, we selected 250 cases of Whole Exome Sequencing Data produced by NextSeq 500, which satisfy all the conditions of Sequencing Depth value of 10 or more and Genotype Quality value of 20 or more. Among them, Intergenic Variant .

(2) Association analysis

Fisher's Exact Test was performed on selected SNPs in the above step, and Association analysis was performed. The correlation analysis was conducted for the whole population (n = 250) and F8 inversion detection group (n = 141), and the comparison between the groups for antibody production was compared with the comparison condition 1 (> 0.60 BU group 1 2) and group 3 (case group) or comparison condition 2 (> 5.00 BU; group 1 and group 2 (control) vs. group 3 (case group)). Variants satisfying P &lt; 0.05 were selected first, with an OR value of &gt; 5 or < 0.2 for the antibody-producing group of the whole population (n = 250) Only SNPs with repeated statistical significance were selected. SNPs with an OR value> 1 among the selected SNPs have a high risk of producing antibodies to the hemophilia A treatment agent. Conversely, SNPs with an OR value <1 have a low risk of antibody production to the hemophilia A treatment agent . SNPs with an OR value> 5 or <0.2 can be considered to have a statistically strong association with the antibody production response.

(3) Variant Annotation

In this step, the mutation of the gene region related to the Immune System was selected for the SNP selected in the association analysis step, and the HGVS (Human Genome Variation Society) information of the corresponding SNP was confirmed.

Example 3: Association analysis of selected SNPs

Table 4 shows the variation of the OR of the antibody-producing group with respect to all 250 patients in which the OR value was > 5 or < 0.2 and P < 0.05.

A list of SNPs with an OR value> 5 or <0.2 for the antibody-producing group in the whole population (n = 250) Genotype Frequency of mutation Phenotype Chr Position (hG19) Gene rsID SEQ ID NO: Reference Opposition Control group Case group OR P > 0.60 BU One 12183245 TNFRSF8 rs2486316 One G T 0.005 0.032 7.036 0.036 > 0.60 BU One 43812255 MPL rs190983971 58 A G 0.005 0.032 7.036 0.036 > 0.60 BU One 43812376 MPL rs117064599 59 A G 0.005 0.032 7.036 0.036 > 0.60 BU One 183559841 NCF2 rs12091406 60 G A 0.009 0.047 5.280 0.011 > 0.60 BU One 207753547 CR1 rs117447527 61 C A 0.005 0.032 7.036 0.036 > 0.60 BU 2 29420550 ALK rs3738869 62 T C 0.005 0.039 8.663 0.011 > 0.60 BU 2 30144064 ALK rs200662898 63 C T 0.005 0.039 8.663 0.011 > 0.60 BU 3 3118142 IL5RA rs17878995 64 G T 0.009 0.049 5.548 0.014 > 0.60 BU 3 57131759 IL17RD rs140930246 65 T C 0.005 0.042 9.485 0.006 > 0.60 BU 3 187009180 MASP1 rs75953973 66 G A 0.005 0.032 7.036 0.036 > 0.60 BU 7 22771296 IL6 rs13306436 9 G A 0.014 0.085 6.605 0.000 > 0.60 BU 8 6727985 DEFB1 rs149045171 67 G A 0.005 0.035 7.847 0.020 > 0.60 BU 11 65427568 RELA rs732072 68 G A 0.009 0.046 5.133 0.012 > 0.60 BU 18 67534642 CD226 rs72481820 69 C T 0.005 0.035 7.847 0.020 > 0.60 BU 21 45712167 AIRE rs78245838 70 T C 0.005 0.042 9.485 0.006 > 0.60 BU 21 46308630 ITGB2 rs117884186 71 G C 0.005 0.039 8.663 0.011 > 0.60 BU 3 52259487 TLR9 - 85 C G 0.005 0.039 8.663 0.011 > 0.60 BU 19 55333193 KIR3DL1 - 86 C A 0.005 0.041 8.153 0.011 > 0.60 BU One 32741561 LCK rs139568272 72 G A 0.056 0.011 0.182 0.004 > 0.60 BU One 161642982 FCGR2B rs2298022 73 G A 0.042 0.007 0.163 0.008 > 0.60 BU 3 9965745 IL17RC rs13097123 74 C T 0.032 0.004 0.106 0.018 > 0.60 BU 3 10084828 FANCD2 rs35625434 75 G A 0.028 0.004 0.124 0.034 > 0.60 BU 6 74524861 CD109 rs185470785 76 A G 0.028 0.004 0.125 0.034 > 0.60 BU 8 22385932 PPP3CC rs61332891 77 A T 0.028 0.004 0.124 0.034 > 0.60 BU 10 97607266 ENTPD1 rs3793744 78 G A 0.028 0.004 0.124 0.034 > 0.60 BU 11 60201102 MS4A5 rs17154981 79 G A 0.051 0.011 0.199 0.008 > 0.60 BU 14 24842563 NFATC4 rs77893724 80 A C 0.028 0.004 0.124 0.034 > 0.60 BU 14 24844669 NFATC4 rs55851358 81 T C 0.037 0.007 0.187 0.015 > 0.60 BU 16 85942721 IRF8 rs11545564 82 A G 0.032 0.004 0.106 0.018 > 0.60 BU 17 32685355 CCL13 rs139142355 83 G T 0.028 0.004 0.124 0.034 > 0.60 BU 21 45648905 ICOSLG rs56292769 84 G C 0.037 0.007 0.184 0.015

Table 5 lists the mutations observed repeated statistical significance for the antibody-producing group and the high-antibody-producing group for all 250 patients. The risk-increasing marker (OR> 1) was 19, the decreasing marker (OR <1) were 23.

SNPs with repeated statistical significance for the antibody-producing group and the high-antibody-producing group in the whole group (n = 250) Genotype Frequency of mutation Phenotype Chr Position (hG19) Gene rsID SEQ ID NO: Reference Opposition Control group Case group OR P > 0.60 BU One 12183245 TNFRSF8 rs2486316 One G T 0.005 0.032 7.036 0.036 > 0.60 BU One 12198297 TNFRSF8 rs476488 2 T C 0.375 0.468 1.468 0.041 > 0.60 BU One 181026031 MR1 rs55700376 3 G A 0.019 0.063 3.553 0.011 > 0.60 BU 3 172223690 TNFSF10 rs1131568 4 C T 0.370 0.479 1.562 0.016 > 0.60 BU 3 172227199 TNFSF10 rs3815496 5 T C 0.324 0.472 1.862 0.001 > 0.60 BU 4 145040999 GYPA rs62334652 6 T C 0.349 0.460 1.591 0.014 > 0.60 BU 4 145041750 GYPA rs45468202 7 A C 0.255 0.351 1.584 0.022 > 0.60 BU 5 40964985 C7 rs3805220 8 G A 0.046 0.099 2.253 0.034 > 0.60 BU 7 22771296 IL6 rs13306436 9 G A 0.014 0.085 6.605 0.000 > 0.60 BU 8 86388228 CA2 rs2307075 10 A C 0.322 0.434 1.610 0.013 > 0.60 BU 8 86389403 CA2 rs703 11 T C 0.343 0.468 1.690 0.005 > 0.60 BU 11 36514245 TRAF6 rs16928973 12 C T 0.051 0.113 2.367 0.012 > 0.60 BU 11 60776484 CD6 rs2074226 13 C T 0.420 0.515 1.465 0.040 > 0.60 BU 12 9747288 KLRB1 rs10844140 14 C T 0.407 0.525 1.605 0.010 > 0.60 BU 16 27356359 IL4R rs2074572 15 C T 0.366 0.472 1.549 0.016 > 0.60 BU 17 73753661 ITGB4 rs9367 16 T C 0.407 0.507 1.497 0.026 > 0.60 BU 19 14092911 RFX1 rs3816553 17 G A 0.074 0.130 1.872 0.048 > 0.60 BU 19 18171886 IL12RB1 rs383483 18 A G 0.333 0.426 1.485 0.037 > 0.60 BU 17 26864392 FOXN1 - 87 A C 0.009 0.039 4.311 0.034 > 5.00 BU One 12183245 TNFRSF8 rs2486316 One G T 0.008 0.055 7.116 0.002 > 5.00 BU One 12198297 TNFRSF8 rs476488 2 T C 0.401 0.508 1.544 0.034 > 5.00 BU One 181026031 MR1 rs55700376 3 G A 0.032 0.078 2.528 0.033 > 5.00 BU 3 172223690 TNFSF10 rs1131568 4 C T 0.403 0.516 1.575 0.026 > 5.00 BU 3 172227199 TNFSF10 rs3815496 5 T C 0.381 0.484 1.526 0.042 > 5.00 BU 4 145040999 GYPA rs62334652 6 T C 0.376 0.516 1.766 0.007 > 5.00 BU 4 145041750 GYPA rs45468202 7 A C 0.273 0.414 1.882 0.003 > 5.00 BU 5 40964985 C7 rs3805220 8 G A 0.062 0.117 2.014 0.042 > 5.00 BU 7 22771296 IL6 rs13306436 9 G A 0.035 0.109 3.372 0.002 > 5.00 BU 8 86388228 CA2 rs2307075 10 A C 0.356 0.468 1.587 0.029 > 5.00 BU 8 86389403 CA2 rs703 11 T C 0.379 0.516 1.744 0.008 > 5.00 BU 11 36514245 TRAF6 rs16928973 12 C T 0.070 0.133 2.038 0.035 > 5.00 BU 11 60776484 CD6 rs2074226 13 C T 0.439 0.571 1.705 0.011 > 5.00 BU 12 9747288 KLRB1 rs10844140 14 C T 0.446 0.555 1.546 0.036 > 5.00 BU 16 27356359 IL4R rs2074572 15 C T 0.395 0.516 1.629 0.020 > 5.00 BU 17 73753661 ITGB4 rs9367 16 T C 0.437 0.539 1.508 0.045 > 5.00 BU 19 14092911 RFX1 rs3816553 17 G A 0.089 0.156 1.902 0.037 > 5.00 BU 19 18171886 IL12RB1 rs383483 18 A G 0.347 0.500 1.884 0.003 > 5.00 BU 17 26864392 FOXN1 - 87 A C 0.011 0.070 6.958 0.001 > 0.60 BU One 158224282 CD1A rs366316 27 A G 0.324 0.232 0.632 0.023 > 0.60 BU 2 202065462 CASP10 rs138067550 28 C T 0.056 0.021 0.367 0.040 > 0.60 BU 3 46479668 LTF rs3213479 29 G A 0.440 0.352 0.692 0.047 > 0.60 BU 3 46491267 LTF rs17141154 30 G A 0.272 0.167 0.536 0.005 > 0.60 BU 3 46491290 LTF rs2269443 31 G A 0.304 0.211 0.612 0.019 > 0.60 BU 5 40928792 C7 rs2459467 32 A T 0.556 0.439 0.627 0.010 > 0.60 BU 5 66478626 CD180 rs2230525 33 T C 0.282 0.204 0.652 0.039 > 0.60 BU 5 133477711 TCF7 rs6876997 34 A G 0.083 0.039 0.443 0.043 > 0.60 BU 5 169468213 DOCK2 rs149956505 35 A T 0.065 0.018 0.259 0.006 > 0.60 BU 10 33201128 ITGB1 rs2504020 36 C G 0.120 0.060 0.465 0.019 > 0.60 BU 10 33211440 ITGB1 rs2252752 37 T C 0.116 0.063 0.517 0.045 > 0.60 BU 11 59836887 MS4A3 rs2298584 38 A G 0.515 0.363 0.537 0.001 > 0.60 BU 12 66591582 IRAK3 rs4762088 39 G C 0.153 0.088 0.535 0.029 > 0.60 BU 16 27364158 IL4R rs3024610 40 C T 0.441 0.298 0.537 0.001 > 0.60 BU 18 2917223 LPIN2 rs1985 41 T A 0.468 0.338 0.581 0.004 > 0.60 BU 18 2934536 LPIN2 rs3765622 42 C G 0.472 0.345 0.589 0.004 > 0.60 BU 18 2937646 LPIN2 rs3826637 43 G A 0.468 0.338 0.581 0.004 > 0.60 BU 18 2954439 LPIN2 rs7226624 44 T C 0.472 0.363 0.636 0.015 > 0.60 BU 19 1623850 TCF3 rs2240592 45 G C 0.338 0.254 0.665 0.042 > 0.60 BU 19 7150418 INSR rs2252673 46 C G 0.285 0.193 0.599 0.016 > 0.60 BU 19 10469919 TYK2 rs55882956 47 G A 0.088 0.025 0.262 0.001 > 0.60 BU 19 54848512 LILRA4 rs56374127 48 C T 0.277 0.164 0.512 0.003 > 0.60 BU 22 37580334 C1QTNF6 rs229525 49 A G 0.421 0.328 0.669 0.028 > 5.00 BU One 158224282 CD1A rs366316 27 A G 0.301 0.188 0.536 0.013 > 5.00 BU 2 202065462 CASP10 rs138067550 28 C T 0.046 0.008 0.164 0.039 > 5.00 BU 3 46479668 LTF rs3213479 29 G A 0.425 0.289 0.551 0.005 > 5.00 BU 3 46491267 LTF rs17141154 30 G A 0.246 0.112 0.387 0.001 > 5.00 BU 3 46491290 LTF rs2269443 31 G A 0.279 0.172 0.537 0.015 > 5.00 BU 5 40928792 C7 rs2459467 32 A T 0.519 0.405 0.630 0.027 > 5.00 BU 5 66478626 CD180 rs2230525 33 T C 0.274 0.133 0.405 0.001 > 5.00 BU 5 133477711 TCF7 rs6876997 34 A G 0.073 0.016 0.203 0.011 > 5.00 BU 5 169468213 DOCK2 rs149956505 35 A T 0.048 0.008 0.155 0.045 > 5.00 BU 10 33201128 ITGB1 rs2504020 36 C G 0.105 0.031 0.275 0.008 > 5.00 BU 10 33211440 ITGB1 rs2252752 37 T C 0.105 0.031 0.275 0.008 > 5.00 BU 11 59836887 MS4A3 rs2298584 38 A G 0.457 0.348 0.632 0.036 > 5.00 BU 12 66591582 IRAK3 rs4762088 39 G C 0.134 0.063 0.429 0.031 > 5.00 BU 16 27364158 IL4R rs3024610 40 C T 0.388 0.275 0.599 0.024 > 5.00 BU 18 2917223 LPIN2 rs1985 41 T A 0.422 0.313 0.623 0.032 > 5.00 BU 18 2934536 LPIN2 rs3765622 42 C G 0.427 0.320 0.631 0.032 > 5.00 BU 18 2937646 LPIN2 rs3826637 43 G A 0.422 0.313 0.623 0.032 > 5.00 BU 18 2954439 LPIN2 rs7226624 44 T C 0.438 0.328 0.626 0.025 > 5.00 BU 19 1623850 TCF3 rs2240592 45 G C 0.315 0.219 0.610 0.037 > 5.00 BU 19 7150418 INSR rs2252673 46 C G 0.255 0.167 0.583 0.044 > 5.00 BU 19 10469919 TYK2 rs55882956 47 G A 0.067 0.008 0.109 0.004 > 5.00 BU 19 54848512 LILRA4 rs56374127 48 C T 0.237 0.147 0.553 0.043 > 5.00 BU 22 37580334 C1QTNF6 rs229525 49 A G 0.393 0.297 0.654 0.050

The results are shown in Table 6, and the risk-increasing markers (OR> 1) were 9, and the decrease was statistically significant for the F8 inversion detected in 141 patients with antibody-producing group and high- The number of markers (OR <1) was 9.

SNPs with repeated statistical significance for the antibody-producing group and the high-antibody-producing group in the F8 inversion detection group (n = 141) Genotype Frequency of mutation Phenotype Chr Position (hG19) Gene rsID SEQ ID NO: Reference Opposition Control group Case group OR P > 0.60 BU 2 242793433 PDCD1 rs2227982 19 G A 0.389 0.506 1.611 0.050 > 0.60 BU 3 148890137 HPS3 rs73019023 20 A G 0.041 0.123 3.291 0.012 > 0.60 BU 5 40945397 C7 rs75345202 21 G A 0.056 0.136 2.671 0.023 > 0.60 BU 5 40964985 C7 rs3805220 8 G A 0.056 0.130 2.532 0.036 > 0.60 BU 5 52367666 ITGA2 rs2287872 22 T A 0.397 0.531 1.720 0.021 > 0.60 BU 5 147493872 SPINK5 rs3815734 23 T C 0.105 0.207 2.224 0.026 > 0.60 BU 16 57418987 CX3CL1 rs2239354 24 G A 0.025 0.117 5.162 0.004 > 0.60 BU 18 60028762 TNFRSF11A rs8099222 25 G A 0.082 0.184 2.529 0.018 > 0.60 BU 20 1896244 SIRPA rs6136376 26 G A 0.350 0.500 1.857 0.012 > 5.00 BU 2 242793433 PDCD1 rs2227982 19 G A 0.429 0.583 1.862 0.048 > 5.00 BU 3 148890137 HPS3 rs73019023 20 A G 0.067 0.182 3.096 0.026 > 5.00 BU 5 40945397 C7 rs75345202 21 G A 0.079 0.208 3.061 0.011 > 5.00 BU 5 40964985 C7 rs3805220 8 G A 0.075 0.208 3.246 0.010 > 5.00 BU 5 52367666 ITGA2 rs2287872 22 T A 0.446 0.604 1.897 0.048 > 5.00 BU 5 147493872 SPINK5 rs3815734 23 T C 0.136 0.283 2.503 0.020 > 5.00 BU 16 57418987 CX3CL1 rs2239354 24 G A 0.062 0.146 2.585 0.050 > 5.00 BU 18 60028762 TNFRSF11A rs8099222 25 G A 0.101 0.326 4.313 0.000 > 5.00 BU 20 1896244 SIRPA rs6136376 26 G A 0.407 0.565 1.893 0.043 > 0.60 BU One 161569893 FCGR2C rs144426992 50 A G 0.294 0.184 0.541 0.029 > 0.60 BU 3 46491290 LTF rs2269443 31 G A 0.331 0.215 0.555 0.026 > 0.60 BU 4 77134870 SCARB2 rs1465922 51 G A 0.381 0.263 0.578 0.035 > 0.60 BU 5 52366138 ITGA2 rs2112290 52 G A 0.540 0.414 0.602 0.038 > 0.60 BU 12 6344600 CD9 rs2072372 53 C A 0.524 0.395 0.594 0.028 > 0.60 BU 12 9747361 KLRB1 rs10844141 54 A G 0.548 0.401 0.554 0.015 > 0.60 BU 18 77156103 NFATC1 rs71359461 55 G C 0.387 0.234 0.484 0.005 > 0.60 BU 19 52132325 SIGLEC5 rs2305372 56 A T 0.468 0.340 0.584 0.025 > 0.60 BU 20 1895889 SIRPA rs1057114 57 G C 0.548 0.414 0.583 0.028 > 5.00 BU One 161569893 FCGR2C rs144426992 50 A G 0.254 0.125 0.419 0.050 > 5.00 BU 3 46491290 LTF rs2269443 31 G A 0.291 0.146 0.417 0.040 > 5.00 BU 4 77134870 SCARB2 rs1465922 51 G A 0.338 0.196 0.478 0.048 > 5.00 BU 5 52366138 ITGA2 rs2112290 52 G A 0.504 0.292 0.405 0.006 > 5.00 BU 12 6344600 CD9 rs2072372 53 C A 0.479 0.313 0.494 0.032 > 5.00 BU 12 9747361 KLRB1 rs10844141 54 A G 0.492 0.333 0.517 0.048 > 5.00 BU 18 77156103 NFATC1 rs71359461 55 G C 0.333 0.146 0.342 0.007 > 5.00 BU 19 52132325 SIGLEC5 rs2305372 56 A T 0.425 0.250 0.451 0.020 > 5.00 BU 20 1895889 SIRPA rs1057114 57 G C 0.500 0.333 0.500 0.033

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

<110> Green Cross Genome Corporation <120> Markers For Predicting Effect of Therapeutic Agent Against          Hemophilia and Use Thereof <130> P16-B131 <160> 87 <170> KoPatentin 3.0 <210> 1 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> K = G or T <400> 1 caaaatgctg tgattacagg tgtgagccaa tgtgcctggc cagcttttcc actttgatct 60 tttgaaacac aaatctgact gtcttagctc ccacggtgtt kcctcgctgg aagccaccag 120 tccctgctgg ttaactcttt cctggagaca ccagcctcct tggcgctggt tctcagatgt 180 tacgtcccct ctgcagatat g 201 <210> 2 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y = C or T <400> 2 agagatgaaa aaaaaaaggg gcctcccagt tcagagactg gtggggaggt tgggggtacc 60 ctgcagcagc acccattccc gtcccacagc agctgaggag yggtgcgtcg gtgacagaac 120 ccgtcgcgga agagcgaggg ttaatgagcc agccactgat ggagacctgc cacagcgtgg 180 gggcagccta cctggagagc c 201 <210> 3 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> R = A or G <400> 3 cctttggctt taatcattac agcctcagat aaaggtacct tcagcccggg cacagtggct 60 cacgcctgta atcctagcac tttgggaggc tgaggcaggc rgattccctg agctcaggag 120 ttcaagacca gcctgggcaa cacggtgaaa ccctgtctct actaaaatac aaataattag 180 ctgggcatgg tggcatgtgc c 201 <210> 4 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y = C or T <400> 4 cactgcaacc tctgcctccc gggttcaaac aattctcttg cctcagcctc tcaagtagct 60 gggactacag gcatgtgcca acacacccag ctaatttttg yactttcagt agagatgggg 120 tttcactatg ttggtcacta tggtcttgat ctcttgatct cgtgatctac ccaccttggg 180 ttcccaaaat gttgggatta c 201 <210> 5 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y = C or T <400> 5 ttttgttgct tttctaaaag agaaatgata aagggtcatc aacacttgcc aaactagttc 60 tccaatacct tgctcttcat aggtgttgtc aatcagccat ygtaagcata ctgggcttcc 120 aaagttgatt ctatgagttt tcataaaaat aacattttga tattatgatc aacctacctc 180 agcttatgat caacttgaaa a 201 <210> 6 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y = C or T <400> 6 taggagtggc tgcatatgtg tcccgtttgt gcgtatctgc atataagaga agttgagaaa 60 gggcacaaga atgaggtgac tgcgtggaca tagagcgtat yatgaaagac aaactctccc 120 acatccctcc cagcagcggc tccagaattt cttcttctcc cctgaaaact aacaagagag 180 actgccacca gattactctt a 201 <210> 7 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> M = A or C <400> 7 ctgtgatgag atgtaactct ttgtgactga agaagaagtt gaagtgtgca ttgccacctc 60 agtggtactt aatgctgata tgctcacaat ttctgtataa matagaagtt gagaaaggga 120 ttaagaacga ggtgactgag cggaccataa agcatattgc aaaagacaaa ttccctcaca 180 tccctccagt ccctgagcta a 201 <210> 8 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 8 acttgcaatg aaggatactc tcttattgga aacccagtgg ccagatgtgg agaagattta 60 cggtggcttg ttggggaaat gcattgtcag agtgagtggc rtcagttgta tataatttaa 120 gatgagaaaa ttacgtggaa gagaatgaat caagataaat aacactcacc atatggccca 180 tgtgttggcc ttcctttcct g 201 <210> 9 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 9 gggcacctca gattgttgtt gttaatgggc attccttctt ctggtcagaa acctgtccac 60 tgggcacaga acttatgttg ttctctatgg agaactaaaa rtatgagcgt taggacacta 120 ttttaattat ttttaattta ttaatattta aatatgtgaa gctgagttaa tttatgtaag 180 tcatatttat atttttaaga a 201 <210> 10 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> M (A or C) <400> 10 ttttcaatac tccattggtt ttagaaattt cttttgatca aattgcacaga tctcaatgac 60 atgtggtgtt tggatgagca gttagtaaag gtagaatatt mcttaaatat ttaattaacg 120 tttctttcaa caaagttgat cctaatgcta gcataattct aagacttgta tttatttaat 180 cttgctcccc catcatttgg a 201 <210> 11 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 11 ctctgtcaat gtgatagttt gaagctgcgt atttgccttg ttctagggca agagtgctga 60 cttcactaac ttcgatcctc gtggcctcct tcctgaatcc ytggattact ggacctaccc 120 aggctcactg accacccctc ctcttctgga atgtgtgacc tggattgtgc tcaaggaacc 180 catcagcgtc agcagcgagc a 201 <210> 12 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 12 ctgtagggca gtctagatca taatgattag gcatctgaaa tccaaaacaa aggctgaaaa 60 atctcttcct tgcatatcat tttctagttt gatgcgatca yatttaaact gtattgtcaa 120 tggctgcttt gtaccacatt ggcagttgaa taggtgcagc ataaaccatc acccacaaaa 180 attaaaagaa catctacttt c 201 <210> 13 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 13 gcctggagtt agtgccggag aggtggtccc tgcccttgag atgtcacagt tcaggagggg 60 agatgagact gagacaaaca taattcagga gagaaagtgg yaaatgcccc caagaaaggc 120 agtcaggtgg agtctagagg cagcagacca tgcggagctg taggctaagg ctgaccacct 180 cccggaagag gtgacattaa a 201 <210> 14 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 14 tacatgagga actcgcaaga gagtgaaatc catagaagtg gcctaagcaa gatgctttta 60 tgctttttag accaagaatg ataaatttga aaagaaatga yaggatgaag aaaagctggc 120 taggggcaat aaattttcta gaggagtcac taggagatat attggttgcg gggggctgta 180 aaataggtga aagataggtg t 201 <210> 15 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 15 ggtcagtgcg gataactata cactggacct gtgggctggg cagcagctgc tgtggaaggg 60 ctccttcaag cccagcgagc atggtgagca gggcggagtg yggcaggggt ggctgggtgt 120 gttcccacag ctgcctgggc tgagggtggg gtgggcaggg gaggaggtgg ggtcatagca 180 acagcaggag gaagccgcct g 201 <210> 16 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 16 ggagtttgtg agccggacac tgaccaccag cggaaccctt agcacccaca tggaccaaca 60 gttcttccaa acttgaccgc accctgcccc acccccgcca ygtcccacta ggcgtcctcc 120 cgactcctct cccggagcct cctcagctac tccatccttg cacccctggg ggcccagccc 180 acccgcatgc acagagcagg g 201 <210> 17 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 17 ggactttgga ggagcaagtg ggcttggggg gcggggcagg gagccccaaa tatactatgg 60 ggatacattg ctgacctcgg gagacagcag gggggagggc rgccaacagg accaccactt 120 acctctgggg gcgagtgcac ctgctgggta ccatggaccg tcagggagac ctggccacct 180 ttgctgcctg gggctgcgct c 201 <210> 18 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 18 gtctttaaaa tgggaagggg tatggagcac tggttttgga gtccttacct tgatgaattg 60 ggagagaata aaatgagcta atgcgtaacc cttgtccagc rtgtgcaccc aataaaaaga 120 cttggaatta gaaaaggcat ccttacctcc ttccctccag ggaactcaat ggcggagctg 180 gcacagggtg tgggcagcgg c 201 <210> 19 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 19 atactccgtc tgctcaggga cacagggcac ggggggctcc ggggtcttct ctcgccactg 60 gaaatccagc tccccatagt ccacagagaa cacaggcacg rctgaggggt cctccttctt 120 tgaggagaaa gggagaggga gtcagcccca cggcccaccg caggaaggag gcccgcggct 180 ccacagcctg gctgcagtac c 201 <210> 20 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 20 gcatttgaga ctgaatttct aaaaattgaa tgccaaagta caagtagagg agttttttat 60 tttatatatc acacacacac acacacacac acacacacac rcacacatat atgatacaaa 120 tgctttcagg ctgcttacct taccgtgtag tggtaactat tcacttctta atttatgacc 180 tcaatcaatt taattgtcta g 201 <210> 21 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 21 taggtgaaaa taaataatga ttttaattat gaattttaca atagtacttg gtcttatgta 60 aaacatacgt cgacagaaca cacatcatct agtcggaagc rctccttttt tagatcttca 120 tcatcttctt cacgcagtta tacttcacat accaatgaaa tccataaagg aaaggttagt 180 ataaaatgtc agttaacttt t 201 <210> 22 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> W (A or T) <400> 22 aagagaggct aaaagcagaa ttaggatgga agaaaagaag cgagagtctt gattattcac 60 aacaaagata ttctacataa ttgagagctg actgtgctct waattgtaaa cacatttgga 120 atcattttct tgttttaatg ttgctatgct ctaataaact gactctgtct ccccctgtat 180 gtttgtgtgt gtgtatgttt a 201 <210> 23 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 23 gtttaatata aattgaacac tataactgcc agaaaaaaaa ttctcaggtc attttctctc 60 tctttttttt ttttctataa agcacttagt aggaacccag yaaactaatt tcccagaaga 120 tactcaagct ttctcctttt cttttccttt taggagacat gcgatgaatt tcggagactt 180 ttgcaaaatg gaaaactttt c 201 <210> 24 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 24 ttttccaacc ctcatcacca acgtctgtgc cattttgtat tttactaata aaatttaaaa 60 gtcttgtgaa tcaatatggg tgatttctct ggggacaagt rcagcaatca gggcttcgga 120 ggggaaggag tggggcaggg ctgggggcaa gacccctttc agccccggag acccctatct 180 ccagttcacc tggggcagaa a 201 <210> 25 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 25 aacaccgtaa tggccctggc tgctctcccg tagagtgaaa ccggactgag cagggctggc 60 tgagattgtg gatctcaccc tttggaatgt gagtgctcgc rgacaaaggg cagccaaggg 120 tggggactgt cacttctgct atcccagacc aacatttttg attctgaggt ttgatttacc 180 gggatttgtt tttcaactgc g 201 <210> 26 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 26 taactcaata ataacaccct ccattctttg agcaatgatc aggtgtggtg gtaggtgctc 60 ctttccatga attgatgtct ccccctttta caaataaggg ragttgaggc ctgaagacgt 120 catgctattt gctgaaggct ccacagctgg taaagggtgg gaacatctgc atccttcatc 180 tgcctggctg tgtagccctg g 201 <210> 27 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 27 ttccatagca gttgaattag gggaaggtga ataagttgga ggttggtgac aaggagagaa 60 gctggaacag agaggagagt cagaaccaga gggaaatgag rgactgagta ggcatctcag 120 ggtttttgaa ggagtggatt ttctttgttg cagtcagggg aggtttgtct gttggctgca 180 gaaagaagtc agaatagaga t 201 <210> 28 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 28 atttgaggga atctcagctt tgggaggcca ctttgtaagt gatcttaaga gattaagtac 60 atccttaatt aaaagaaatg gtaacacctc cgcctgcttg ytgtgagttt attcatttga 120 attccccatc tcccccaatg ggaaataaca cagaattgca attatttata aaatcttaaa 180 ttggcttctt ttctctagga a 201 <210> 29 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 29 tggaataagc aaaacttgtc cgggcagtca gatccatttc tcccaaattt agcctgcgac 60 aaaagggcag acagtgagta gctaaggaaa agaggaattt rtgggttaaa ggaggcaaag 120 gggtatttct tccacccaag aacagccctg agaaaaccat ggggcagaag gagctgagtt 180 ctggagtcat tccacctcct t 201 <210> 30 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 30 gactattctt ttagagacac ctgacctaat ccacagaaga cactcacacc tgcatcaaac 60 ctccacgatg accccacagt gtctgggaaa aggagtgacc rccacaaagt agggccacct 120 gctgggaagt agaagaccac accaggcgga cctcaggacc ctcctgggct cactcacttt 180 tcctcaagtt ctggatggca g 201 <210> 31 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 31 acctaatcca cagaagacac tcacacctgc atcaaacctc cacgatgacc ccacagtgtc 60 tgggaaaagg agtgaccgcc acaaagtagg gccacctgct rggaagtaga agaccacacc 120 aggcggacct caggaccctc ctgggctcac tcacttttcc tcaagttctg gatggcagtg 180 aagtagccgg agccaaggta c 201 <210> 32 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> W (A or T) <400> 32 tattcatttt ggtgggattt ataggagagt tccaaagttt ttcaaggtga ggactttttg 60 ggttgtgtgt aaaaatcatt aaatttaaaa aatgttttag waattcttta gtctaatttt 120 gacattgcac tttgaatctg tgtatccctc caacatattt tctctttcca tatttttata 180 aaagacaatg ataacaaaag c 201 <210> 33 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 33 actcacagga gtaagaagct cagaaaagca cagtgagtcc ctgcccagtt ccaggaagca 60 atctgaaaag tctggtctgg tcaccagcag atgacagttc yttcacttag agcaattttg 120 ctaagcacac ttatttgctt tctctggaaa ccttcagcac taaatgtgtt ggtatttcca 180 cctgagaagg tatttaactg c 201 <210> 34 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 34 aaggggacag attcaaaagc tacttgggac ctatgtagta accatctgaa tggcaatcta 60 tgtagatttt tgcagtgtgt ctgtgtagtg tgctgggagc rggtatttgt atttatctct 120 gtgtacttat gtctaggcca gtaggataga gtatctatct gttcacctgt gtcctcaagt 180 tatgattat gcaggggcct c 201 <210> 35 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> W (A or T) <400> 35 tgcctacacg ctccttctcc acacctggct tctcaaggta cagtcacttt gggtgaaggg 60 catttatgct gggatcctga cagggcctca cagcaagttg wagagtgaaa aaagaaaaaa 120 aagaaaaaga atccaaatat tttgttttgc cctgtgtctc caaactcttg agctggcagg 180 atactttttg cactgcttcc c 201 <210> 36 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> S (C or G) <400> 36 tctaccaaca cgcccttcat tgcacctgaa gaaacatgcc aaaattgcaa tcacacaaaa 60 caaaaataat ttaagaggtt ccaagatggc caaacaggaa sagctctagt ctacagctcc 120 cagcgtgagc gacacagaag acaggtggtt tctgcatttc caactgaggt actgggttca 180 tctcactggg gcttgctgga c 201 <210> 37 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 37 agcaactttt atctttaaat gccagttccc atgtttcctc ctaactaact ttaggagtca 60 aacctagaaa ccagttttat aatatgtgat taaaaattaa yataaaccac ttttgtaaaa 120 ttcggtttgc ctatctggtg gcattttccc acatttcaat tcagacatga tttgcctcta 180 acaatcaaaa atgtttagaa t 201 <210> 38 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 38 agcttcttcc ccctgtagct acttcacatg cgttgacata gaaatgtgac acatctttct 60 cattagagaa aatgttttca tgcttttaga ttgagatact rtctcttcta cagtgttata 120 tgtgtctctg tctttaattt atggttcact ttgttccact ttacgggagg gagatgttgt 180 agtctggttt cctgctgaca t 201 <210> 39 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> S (C or G) <400> 39 atgtgatgca aactcagaac tatatgcccc ggtcaatgtt agagacccta aaaccccagc 60 tcatttggca tctgttcatt catattccag ctgaatctca saaatgtgct taagggaaag 120 catctgatat tagcagtttc tcagccaagt cacaggacgg gacatcccag agtcatggcc 180 tgctgcactt ttaaaaggga a 201 <210> 40 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 40 ccattcttcc cctgtggcca gacacttccc ctggctgagt ctctgggctt ttatatcata 60 ggatgcctct aatggcaatc ctgccattag atacacctgc ygtggtgtat ctgccaggta 120 ggcaggctag gctgcagtaa caaacaagcc cacaatttcc atggcttaac actatgggaa 180 tatatttctt gctcacgtaa c 201 <210> 41 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> W (A or T) <400> 41 acaattaact ttggacaact taaaacttat tagtgacatt gctgtctaat aatcaaatac 60 ttcatcatag gctgaacata attattaaaa gagcaaagtt wcccctccct ttcttacttt 120 caaacaaaac caaaagagta gttttcatct ggaaagagca cttacttcag ggaaagactc 180 aatttttaac cagttttatt a 201 <210> 42 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> S (C or G) <400> 42 cttttgtgaa cacctgttta aaaaaaaaat cagaggtaag aatttagtta ttcttcattt 60 acagctctgc aaaacacacg cacgtttgca aacagtaaga stgacaagca ggatttgaat 120 agggttttaa ttttaaaggt gtcaaccagc ctttttactt aggttggttc ccagccatcc 180 atgccaatag aagagaagtt c 201 <210> 43 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 43 aaaaaaaaaa aaaaaaaaaa aaaagtgcga cgggtttcga ctggtgaata caaatacaga 60 ggcagccatc acgttatgtg gaacactgaa ataatttacc rtctaatttg agagtacctg 120 gagccaaatt aaacaaacga ttacctttct tctttgacgg cgagtctact ttagctgccg 180 gtttggattc tgagggcgcc t 201 <210> 44 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 44 tcctttttga cagcttagtt cttcaacatg aaactaaagc ttgtcttgcc aacaacagtc 60 ctctgtacaa tcaaccataa aaaactgcta aacggtccgt ytgggccacg taccagaggc 120 ctgagcacac tgtgtaagga gggtgtaacc catgcaaact tacatattct tcttcagtct 180 cctcaacaaa gaaagcttct c 201 <210> 45 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> S (C or G) <400> 45 aggctggacc tagggaagga cttccccgcc ttcccttgga ggcggcctcg ggtcagagct 60 cagatcttgg ctcagaaatc acagggggcc tgtccactca sggcccaccc cgccatgtgt 120 gttcccaagc ttcgccagga cacagggcct ggcactgcca ctgacgagct gtggggtccc 180 ttctccctcg tgcgactcac c 201 <210> 46 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> S (C or G) <400> 46 cattcatgcc cagatttcat ttcagagtga aggcattgga tttctgaatt ggtgaagcat 60 ctgctctcca gcacagctgc ccgccgcatg caaaaagcca sagaaacccc tgggttctcc 120 gaggcatctg cctggcacgc cgccggccct gcgcggagca ggcaccaggg gtcgcacagg 180 tgagtcatac ctagggtcct c 201 <210> 47 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 47 accgacccag gccccacaca caggccacac accaggtagc tgagggcgct ggccagctgc 60 tgggccacca ccatcttcca agccatgggc acatggcccc rctccctccg cagccacaca 120 tccaggggtc cgtgctccac gtactctgtc accatgatat ctgtaaagac acagctgctc 180 tggggcaacc tgcctgcttc t 201 <210> 48 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 48 gtcctggaga aaagaaggac gggtgagggg ctgccccacc ttgttctgag ctgagacctc 60 cccaccagtc ctctccctgg gaccctcagt ctgtctctgt yttctctgag tctccccctc 120 cccgcccatc ccctgtctct gtctgtctct ccctccctta ggacccccac ccctcatccc 180 ggccatcacc acctgggctc c 201 <210> 49 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 49 ggtgccaggt gaccactggc tgtgcagtga gtgtccccgg ggagagctca ggaacaaatc 60 ctggctccat ccctgacagc tgtgtggccc caggagactg rgtttcccca tctgcaaagt 120 gggaggaata agaaaatatc gacctcacgg gctgttgtgg gcagagacgg acgctaagga 180 tgtcaaggcc tggtcacagc c 201 <210> 50 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 50 ataagcaaaa cttaacttgg atgatttctg gtaaatgctt atgttagaaa taagacaacc 60 ccagccaatc acaagcagcc tactaacata taattaggtg rctagggact ttctaagaag 120 atacctaccc ccaaaaaaca attatgtaat tgaaaaccca tcgattgcct ttattttgct 180 tccacatttt cccaataaat a 201 <210> 51 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 51 cagaggcgtc gaagacccgg gacccttcgg cgccacgccc acgccctccc ggcgcacggt 60 tcgtgcgcgc agctctgggc tccgcggcct ggcgagcgcg rccccgggtg caccgggcgg 120 atggggccgc ggagggacgg gcccggactc ggtttcggtt tccttcgccg ggcagccgtg 180 gcgcccgcct agcgcagctc g 201 <210> 52 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 52 atatggtagt aaatcaagca cagagttgcc ccgagcacat catttatata caggtaaggc 60 ctcaggaaca tcccttttga ctttatttct tcctaaagac ragagagtgg taaaagtcaa 120 aaatggaagc ttatgagtta gaaaatgctc acccttccaa tgaatgcctt tcttatgtaa 180 ttccataaat aagaagcata t 201 <210> 53 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> M (A or C) <400> 53 cgccttggca aagacaccct cctgcgcttt ctccggattg tgtctgcaca cagggtgctg 60 actgcaccag accagtgcaa acattctaat cgtcttctta maatttgttt ctctcatccc 120 catccctgcc ttctcgctgt agttgaactg ctgtggtttg gctgggggcg tggaacagtt 180 tatctcagac atctgcccca a 201 <210> 54 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 54 aagaatgata aatttgaaaa gaaatgacag gatgaagaaa agctggctag gggcaataaa 60 ttttctagag gagtcactag gagatatatt ggttgcgggg rgctgtaaaa taggtgaaag 120 ataggtgttt attaagtatc tttattcagg ttcattgcag catccaattc caggtctctg 180 gcgataagag ctattttctt g 201 <210> 55 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> S (C or G) <400> 55 ggcaccgggg cgcgggcagg gctcggagcc accgcgcagg tcctagggcc gcggccgggc 60 cccgccacgc gcgcacacgc ccctcgatga ctttcctccg sggcgcgcgg cgctgagccc 120 ggggcgaggg ctgtcttccc ggagacccga ccccggcagc gcggggcggc cgcttctcct 180 gtgcctccgc ccgccgctcc a 201 <210> 56 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> W (A or T) <400> 56 tctctacctc tgctcagaga cagacccaga ggcaggtccc agcttcagag aggaggtctt 60 tcctacctat gccgttcctg aagatggtga tggtctgtgg wgcatctggg ataaaaagat 120 ataaacttgg cttcagcagt gaggagtgag atgggcatgg gcccaggagg tacccaaaga 180 ggagccacag aaacccacca a 201 <210> 57 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> S (C or G) <400> 57 tccgtgttgg ttgcagctgg agagacagcc actctgcgct gcactgcgac ctctctgatc 60 cctgtggggc ccatccagtg gttcagagga gctggaccag sccgggaatt aatctacaat 120 caaaaagaag gccacttccc ccgggtaaca actgtttcag acctcacaaa gagaaacaac 180 atggactttt ccatccgcat c 201 <210> 58 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 58 aaatccagga ctacagaccc cacagttctc tcgctgccac ttcaagtcac gaaatgacag 60 cattattcac atccttgtgg aggtgaccac agccccgggt rctgttcaca gctacctggg 120 ctcccctttc tggatccacc aggctggtaa gaactttctt cctcattctt cccacatagt 180 tcccaccccc actgaatctg a 201 <210> 59 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 59 tcccctttct ggatccacca ggctggtaag aactttcttc ctcattcttc ccacatagtt 60 cccaccccca ctgaatctga ccctgtgccc aggatcccca rctctgaccc ttctgaccga 120 tggctctggt ggcacaatgc cttgtgcaca gaaggactta agctgctccc tgctgacatc 180 cctgtagtgc gcctccccac c 201 <210> 60 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 60 gaggagagag gaagtacatc agctgctgcc tgaaggatca gttccccaaa atgcttctgc 60 tctgtcaccc agctcctctt ctctctccca cccctgttct rtggcagctt ctggccaaag 120 ttctgcttct agagccaggg taaccttggc aggtagaggc acctccctgg tgataatgac 180 aggaggcagc caaatgccta g 201 <210> 61 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> M (A or C) <400> 61 atttacaagt gaatttgggg ccttgtgcta gggagaattg ggttctattt ctctacctct 60 gactagctat gaggtcttca ggaagcatag gaaattcccc mataactaac aagtactctg 120 gaactgtcct ttccacagtg tgtcagccgc ctccagaaat cctgcatggt gagcataccc 180 caagccatca ggacaacttt t 201 <210> 62 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 62 ctggtgacaa actccagaac ttcctggttg cttttgctgg ggtatggcat atatccaaga 60 gaaaagattt cccatagcag cactccaaag gacctgggca ygggacagag gacatggaga 120 tggatataga cacacccacc cacattcaca cacacacaca ggcacacaga catacaattt 180 caaccttttt ttcccccatt a 201 <210> 63 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 63 aggctgagaa cggcggctcc cagctgctgc acgctgtcct ggccgccttt tgcgttcctt 60 ttggctcctc caagctcttc tgcccggtct gggcgggaac ygagggcgga ggctgccgtc 120 ttgcgcaccc tcaagctatc tctccgctgc gggaaggctt cggactgtct gcctgctgaa 180 cttctgggcg tgaatcccag c 201 <210> 64 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> K (G or T) <400> 64 acatttccac ttttgatttg gcctacactc atcttggaca ggtcatctag gcacaaaaat 60 gccacaaatt ttacctgata ataagaaccc tctgcctacc kgatgcatgg taagcctgta 120 aaaatgcagt actgaaaccc cactgcaagc acgcaaatgt aaagaacact tacattttac 180 agataagcga gagaattaac a 201 <210> 65 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 65 gaagacgttt ctgtctggtc cgtcgagagt ccttccatca gtggcagaga cagctcggat 60 gagggcacag acgagtcata gatgcctgag tcccgcggca ygtccgaggg gctgccggct 120 ttcaccgtgt gcagcagggg ttgcagggcg gcgctaccgt caagggcagg ccgggcctcc 180 ccgtcttggt ccaggccccc a 201 <210> 66 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 66 ggaaaaatca gccttgtggg tgaacaggag gagagtcaca ttcgccagca ggcagcaccc 60 tgtgcaggca gtgggagaaa tctgagcttc agcagcgtcc rgagcaggat ttgcacgaag 120 tctcccgttt caagttttat gtctcctgga gtcctgggga aggcacccag cccacccaca 180 gctcccttct cagagcccta a 201 <210> 67 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 67 aaaaaaaaaa aaaaaaaaaa accaaaaaaa aaaaaaccaa aatcagtcac ctgttggttt 60 gtggatcaaa caaagtggag gtgacatcac tgaagcctag racatcactc cccttttcca 120 ggcaggcttt gtggttcgac ctgtctcacg ttccaaccca tgcaccctaa ccccctactt 180 ctttacctct tccaattgag 201 <210> 68 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 68 tgggagagta ctgaatgggc accaagattc cagctttact gtgtcttggc cagtgaggga 60 gatgcaggaa aggcgggctg gggagggtga cagagggtgc rggtgtggca ggcaggtttg 120 ccctcacctt ggaaggggtt gttgttggtc tggatgcgct gactgatagc ctgctccagg 180 tcccgcttct tcacacactg g 201 <210> 69 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 69 ttgcatctgt aaattacaaa agaaaaaaat gacagttccg tataaactta ccttctgtgt 60 atcccaggac tctgtaaata gatctcttct ctctctcctt ytccttctgg aatgcatatt 120 caataaagga tataaagata tgatattcca gtactaatat tatttcgaag actctatacc 180 tcactgtacc cctagcccac 201 <210> 70 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 70 ctcggttccc cctctgtgaa aagacatggt cggagccctg gagctccacc cgtgggtttg 60 gggatctgtc acccgctgtc ttgttctgca tgtctctgac yggtggacac acgagcagtg 120 ggacctggag gtgctccagc tgcctgcagg caacagtcca ggaggtgcag ccccgggcag 180 aggagccccg gccccaggag c 201 <210> 71 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> S (C or G) <400> 71 ccgcaccacc ctcgcctctg cggagacccc gcagccggag ctctggagcc ggtccccgct 60 gcaccccagc ggcctcacct cggctctcat ccacatagat saggtagcgg tccatcccgt 120 cctgctgctc cagcgtgtag gccacccagc agccctctga gtccctctcc ttgcaggtcc 180 tgcccttcac ggggttgttc g 201 <210> 72 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 72 gggccagact cactgcgttc tttcgtcgct ttgtccatcc attcattcat tcaggatcgt 60 tttcactgtc ggtccgggac ttcgaccaga accagggaga rgtggtgaaa cattacaaga 120 tccgtaatct ggacaacggt ggcttctaca tctcccctcg aatcactttt cccggcctgc 180 atgaactggt ccgccattac a 201 <210> 73 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 73 aatccaagaa attttcccgt tcggatccca acttctccat cccacaagca aaccacagtc 60 acagtggtga ttaccactgc acaggaaaca taggctacac rctgtactca tccaagcctg 120 tgaccatcac tgtccaaggt atgcggagtc tgccaagatg taaggagggg agaagagggg 180 atggacaagg gctgaggtca c 201 <210> 74 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 74 ctctccctgt actggaatca ggtccagggc cccccaaaac cccggtggca caaaaacctg 60 gtgaggcctc ccccttccca agtccattcc cactgtaggc ygatgcctgt gcaaaggacg 120 cagtgccata tcagagagga tccttgaaga ggactcaccc caagcaaggg aaaattggtg 180 ggggaacttc tgccttcctg g 201 <210> 75 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 75 ctgcaggtaa tttggagct tcgggagaag ttggatctgc agcattgtgt tttgccatca 60 cggttacagg cttcccaagt aaagttgaaa agtaaaggac ragcaaggta aagagctcat 120 cctcacacag gatgtcacaa ttttctgaca tcccactgtc agagttagag cttaatacta 180 ctacaaataa tgatactatt a 201 <210> 76 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 76 caaaatcaag aagcctttga tttagatgtt gctgtaaaag aaaataaaga tgatctcaat 60 catgtggatt tgaatgtgtg tacaaggtaa gtgtctgctt rggtctctct tctttttttc 120 ctttaaaaaa tagacttgaa ggtttaatta tgtatagttg tctatatcaa tctaagagtt 180 atattgaaca aagaattcag t 201 <210> 77 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> W (A or T) <400> 77 tacccaaatt ctttttaaac ctgaaagttt acatttatgc tactttttat agctattctt 60 actaacataa tgataaataa tttttgctta tgcttactta wcatgtgtgt ttatttagat 120 atccacatta ttttagagaa agtacctcat taatcgcgta aattttggat tattttgtct 180 tacttagtca cagagatgct g 201 <210> 78 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 78 ttcacacatg atgctttcaa gtgatttttc cattcaggtt gcaagtaatg aaattctcag 60 ggacccatgc tttcatcctg gatataagaa ggtagtgaac rtaagtgacc tttacaagac 120 cccctgcacc aagagatttg agatgactct tccattccag cagtttgaaa tccagggtat 180 tggaaactat caacaatgcc a 201 <210> 79 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 79 cttaggcata gaatatgggg aatggaaagg ttactaaaag ttcacaagat ccccttattg 60 tgtagatgaa ggcctgaagc ccaaagatgt gaagcaattc rcacaggatc acagagctgc 120 ctcgtggcat taagatgctg tgactaaaat cctgctctcc attctccgca ctcattgctt 180 tgtttgtagt acaggctgga c 201 <210> 80 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> M (A or C) <400> 80 tcaggtgcac cgtatcacag gcaagatggt ggccacggcc agctatgaag ccgtagtcag 60 tggcaccaag gtgttggaga tgactctgct gcctgagaac macatggcgg ccaagtaagt 120 cccatgcaac ttcccctcag tccgcaggct ttgtactagc tttctccact gggcctatgc 180 tagcccactt cttccttttc c 201 <210> 81 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> Y (C or T) <400> 81 tttggagctg ggtgggaggg ctcaagggtt gcttccgtta ctgactccct ctggtttctg 60 ggctggccct gccatttccc ttggtctcag tttgcttccc yataccttga ggacagtttt 120 aaaccctacc tcttccactc atattaagtt aacatgcagt gggctcctct gtgtgtcaga 180 ccttctaggg ccttctgaat t 201 <210> 82 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> &Lt; 223 > R (A or G) <400> 82 ttaaagaagg ggacaaagct gaaccagcca cttggaagac gaggttacgc tgtgctttga 60 ataagagccc agattttgag gaagtgacgg accggtccca rctggacatt tccgagccat 120 acaaagttta ccgaattgtt cctgaggaag agcaaaaatg taactatcct ttatgggcat 180 gaaaccttcc agaagctgcc c 201 <210> 83 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> K (G or T) <400> 83 aaataggatg cattcggttt tgtgattcaa aatgtactat gtgttaagta atattggcta 60 ttatttgact tgttgctggt ttggagttta tttgagtatt kctgatcttt tctaaagcaa 120 ggccttgagc aagtaggttg ctgtctctaa gcccccttcc cttccactat gagctgctgg 180 cagtgggttt gtattcggtt c 201 <210> 84 <211> 201 <212> DNA <213> Homo sapiens <220> <221> variation <101> <223> S (C or G) <400> 84 gctgccccaa gtgctcccct gcagtctggg agtccatgct caagctgcca cctggcctct 60 ccggtggcg tctcacggaa gccctgtcca cgctctgggc sgtgagctcc ggtcaaacgt 120 ggccttggaa gggagatgaa ggagagagag gaacatgaag cacccctttc ccttctgcag 180 gcagacgtcg gttttctcag g 201 <210> 85 <211> 200 <212> DNA <213> Homo sapiens <220> <221> variation <100> <223> S (C or G) <400> 85 ggcttcggct ctgaagtctt caggattcag tgagtgtccc cagggccatg tgggtgacaa 60 cccgtcactg ttgcttgcag ctgccaagcc cacttcttts cccacccctt cccaggatat 120 ccccttcccc aggggactga gagctgttgt cctaccatgc tggggggcag gggcttctcc 180 agagggtctg gcgggcagac 200 <210> 86 <211> 200 <212> DNA <213> Homo sapiens <220> <221> variation <100> <223> M (A or C) <400> 86 tgtagctccc ggagctccta tgacatgtac catctatcca gggagggggg agcccatgaa 60 cgtaggctcc ctgcagtgcg caaggtcaac agaacattcm aggcagattt ccctctgggc 120 cctgccaccc acggagggac ctacagatgc ttcggctctt tccgtcactc tccctacgag 180 tggtcagacc cgagtgaccc 200 <210> 87 <211> 200 <212> DNA <213> Homo sapiens <220> <221> variation <100> <223> M (A or C) <400> 87 gacttggcag ccccgggcag tggtggctcc ggggcactgg gtgacctgca cctcaccacc 60 ctctactctg cctttatgga gctggagccc acgccccccm cggcccctgc aggcccctct 120 gtgtacctca gccccagctc caagcccgtg gccctggcat gagctgtgcc cagcttcgtc 180 agctccagcg tttgcctggt 200

Claims (27)

delete delete delete delete delete Polynucleotides composed of 5 or more consecutive bases derived from the nucleotide sequences of SEQ ID Nos. 65, 69, and 70 and comprising the 101st base of SEQ ID Nos. 65, 69, and 70 as SNPs, or complementary polynucleotides thereof (Factor VIII, F8) as an agent for the treatment of hemophilia.
7. The composition according to claim 6, wherein the agent is a probe.
7. The composition according to claim 6, wherein the agent is a primer.
7. The composition according to claim 6, wherein the preparation is a platemer.
Factor VIII, F8, which comprises the composition of claim 6, as an agent for the treatment of hemophilia.
11. The kit according to claim 10, wherein the kit is a PCR kit, a DNA chip kit or a fluorescent dye kit.
A polynucleotide comprising 5 or more consecutive bases derived from the nucleotide sequences of SEQ ID NOS: 65, 69, and 70 and comprising the 101st base of SEQ ID NOS: 65, 69, and 70 as SNPs, or a complementary polynucleotide Factor VIII, F8, as a therapeutic agent for hemophilia.
(a) polynucleotides composed of 5 or more consecutive bases derived from the nucleotide sequences of SEQ ID NOS: 65, 69, and 70 from the DNA of the separated sample and comprising the 101st base of SEQ ID NOS: 65, 69, Amplifying complementary polynucleotides thereof; And
(b) determining the base of the amplified polymorphic site of step (a). 8. The method according to claim 7, wherein the factor VIII is selected from the group consisting of Factor VIII and Factor VIII.
(a) polynucleotides composed of 5 or more consecutive bases derived from the nucleotide sequences of SEQ ID NOS: 65, 69, and 70 from the DNA of the separated sample and comprising the 101st base of SEQ ID NOS: 65, 69, Hybridizing complementary polynucleotides thereof with a probe; And
(b) determining the base of the hybridized polymorphic site of step (a). 8. The method of claim 7, wherein the factor VIII, F8 is selected from the group consisting of:
The method according to claim 13 or 14,
The polynucleotides derived from the nucleotide sequences of SEQ ID NOS: 65, 69 and 70, or complementary polynucleotides thereof, of the nucleotide sequence determined in the step (b), wherein the polymorphic site is the allele of Table 1, (Factor VIII, F8) is determined to have a high risk of antibody production.
delete delete delete The method according to claim 13 or 14,
The base crystals of step (b) may be analyzed by sequencing, hybridization by microarray, allele specific PCR, dynamic allele-specific hybridization (DASH), PCR extension analysis, PCR- (SSCP), a PCR-restriction fragment length polymorphism (PCR-RFLP), and a TaqMan technique.
delete The polynucleotide of claim 6, wherein the polynucleotide comprises at least 5 consecutive bases comprising the 101st base of SEQ ID NOS: 1, 9, 58-64, 66-68 and 71 and the 100th base of SEQ ID NOS: 85 and 86 as SNPs Wherein the polynucleotide further comprises at least one polynucleotide selected from the set of polynucleotides or complementary polynucleotides thereof.
7. The polynucleotide according to claim 6, wherein the polynucleotide further comprises at least one polynucleotide selected from polynucleotides composed of 5 or more consecutive bases comprising the 101st base of SEQ ID NOS: 72 to 84 as a SNP or a complementary polynucleotide thereof &Lt; / RTI &gt;
7. The polynucleotide according to claim 6, wherein the polynucleotide comprises at least one polynucleotide selected from polynucleotides composed of 5 or more consecutive bases comprising the 101st base of SEQ ID NOS: 2 to 8, 10 to 57 and the 100th base of SEQ ID NO: 87 as SNPs Lt; RTI ID = 0.0 &gt; polynucleotide &lt; / RTI &gt; or a complementary polynucleotide thereof.
15. The polynucleotide according to claim 13 or 14, wherein the polynucleotide comprises the nucleotide sequence 101 of SEQ ID NO: 1, 9, 58 to 64, 66 to 68 and 71, and the nucleotide sequence of SEQ ID NO: 85 and SEQ ID NO: Wherein the polynucleotide further comprises at least one polynucleotide selected from polynucleotides composed of at least two consecutive bases or complementary polynucleotides thereof.
15. The polynucleotide according to claim 13 or 14, wherein the polynucleotide comprises at least one polynucleotide selected from polynucleotides composed of 5 or more consecutive bases comprising the 101st base of SEQ ID NOS: 72 to 84 as SNPs or a complementary polynucleotide Further comprising the step of:
15. The polynucleotide according to claim 13 or 14, wherein the polynucleotide comprises polynucleotides consisting of 5 or more consecutive bases comprising the 101st base of SEQ ID NOS: 2 to 8, 10 to 57 and the 100th base of SEQ ID NO: 87 as SNPs &Lt; / RTI &gt; or a complementary polynucleotide thereof. &Lt; / RTI &gt;
26. The method according to claim 25, wherein the polynucleotide comprises at least one polynucleotide selected from the group consisting of SEQ ID NOS: 72 to 84 or a complementary polynucleotide thereof, wherein the polymorphic site is the allele of Table 2, And judging that the risk is low.

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