KR101646189B1 - Marker for diagnosing intrinsic atopic dermatitis and use thereof - Google Patents

Abstract

One or more embodiments provide SNP markers associated with the invention of endogenous atopic dermatitis, microarrays and kits comprising the markers, and methods of identifying individuals with altered risk of developing endogenous atopic dermatitis using the markers.

Endogenous atopic dermatitis, SNP

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a marker for diagnosing endogenous atopic dermatitis,

One or more embodiments relate to markers for the diagnosis of endogenous atopic dermatitis, compositions comprising the same, microarrays, kits and uses thereof.

Atopic dermatitis (AD) is a common, chronic, inflammatory skin disease that has spread worldwide. In atopic dermatitis, the skin is dry, easily irritated, causes immediate hypersensitivity in allergic reactions, is often thick, usually reddened, often infected, often exudatvie, especially itchy (itchy). Atopic dermatitis is characterized by the ability to form IgE antibodies that cause immediate hypersensitivity when exposed to a specific allergen. Expression of these phenotypes is known to involve multiple genetic factors. Atopy sensitivity genes include several HLA class II molecules, IL-4 receptor proteins, and IgG high affinity receptor proteins.

Atopic dermatitis can be divided into extrinsic type AD and endocrine atopic dermatitis (intrinsic type AD). Exogenous atopic dermatitis is atopic dermatitis associated with an allergic factor with high serum IgE levels and allergen-specific IgE. Endogenous atopic dermatitis is a nonallergic atopic dermatitis that exhibits normal skin IgE levels or skin lesions and distribution patterns similar to extrinsic atopic dermatitis. Patients with endogenous atopic dermatitis account for 10 to 45% of all patients with atopic dermatitis.

As described above, endogenous atopic dermatitis has the same symptoms as allergic atopic dermatitis, but serum IgE level is low to normal range and allergen-specific IgE is not detected. Therefore, preventive management through early diagnosis of high risk through genetic testing It is important.

One embodiment provides a polynucleotide having an association with the onset of an endogenous atopic dermatitis of an individual, and a microarray, composition, and kit comprising the same.

Another embodiment provides a method of identifying a subject having an altered risk of developing an endogenous atopic dermatitis of an individual.

One embodiment is a polynucleotide selected from the group consisting of the nucleotide sequences of SEQ ID NOS: 1 to 9, or a complementary polynucleotide thereof, comprising at least 10 consecutive nucleotides selected from the polynucleotide, including the SNP position of the polynucleotide Wherein the SNP position is a 101 < th > nucleotide, to provide a polynucleotide for diagnosing the risk of endogenous atopic dermatitis in an individual.

The term " intrinsic atopic dermatitis (IAD) "is used by the person skilled in the art in a widely known sense. The endogenous atopic dermatitis may be a chronic or recurrent skin disease, involving itching, dry skin, and / or characteristic eczema. "My impression of the risk of developing atopic dermatitis" may be a relative risk of developing endogenous atopic dermatitis. For example, the risk may be indicative of whether the probability of onset is increased or decreased relative to a normal group. In addition, the risk may be indicative of whether the individual having a particular allele or genotype has an increased or decreased probability of developing atopic dermatitis as compared to an individual having another specific allele or genotype. The entity may be a human. For example, the human being may be a Korean.

"Polynucleotide" may be DNA or RNA. The polynucleotide may also be in single stranded or double stranded form. The polynucleotide may also be a polynucleotide which is not only composed of a natural nucleotide but also a natural nucleotide, an analogue of a natural nucleotide, a sugar, a base or a phosphate site of a natural nucleotide, which is capable of being hybridized to a complementary nucleotide by hydrogen bonding (Scheit, Nucleotide Analogs, John Wiley, New York (1980); Uhlman and Peyman, Chemical Reviews , 90: 543-584 (1990)), and nucleotides selected from the group consisting of . The polynucleotide includes PNA. Further, the polynucleotide may be attached with a detectable label (e.g., Cy3, Cy5 fluorescent substance), for example, in order to facilitate detection of the polynucleotide or the complex to which the polynucleotide is bound in the assay reaction, Terminal or 5 ' end.

The polynucleotide represents a single nucleotide polymorphism at the SNP site. Thus, when one single-stranded polynucleotide is associated with the risk of developing atopic dermatitis, it can be judged that the polynucleotide complementary to the single-stranded polynucleotide is naturally associated with the risk of developing atopic dermatitis. Thus, a polynucleotide according to one embodiment comprises a single-stranded polynucleotide and a polynucleotide having a complementary sequence associated with the risk of developing atopic dermatitis having one specific sequence. For example, the polynucleotide of SEQ ID NO: 1 has the nucleotide at position 101 (SNP position) is "C or T ". In this case, the polynucleotide includes 10 or more consecutive nucleotides selected from the polynucleotide of SEQ ID NO: 1, including the " C or T "nucleotide at the 101st (SNP position) Position "G or A" nucleotides. In this regard, all sequences herein are referred to as the promoter sequence identified by the rs number of the SNP registered in the NCBI (http://www.ncbi.nlm.nih.gov, national center for biotechnology information) GENBANK (flanking sequence).

Single nucleotide polymorphism (SNP) is used in the sense commonly known in the art. A SNP can represent a single nucleotide polymorphism present in a genome within a population. The SNP may have a frequency of a minor allele of the SNP in the population of 1% or more.

The polynucleotide may be 10-100 nucleotides in length. For example, the polynucleotide may be 10 to 50 nucleotides in length, or 10 to 30 nucleotides in length.

The polynucleotide may be a primer or a probe. "Primer" refers to a single-stranded polynucleotide capable of acting as a starting point in polymerization of a nucleotide by a polymerase. For example, the primer can be a single primer that can serve as a starting point for template-directed DNA synthesis under suitable conditions (i.e., the presence of four different nucleoside triphosphates and polymerization enzymes) in a suitable buffer and a suitable temperature. Strand < / RTI > polynucleotide. The appropriate length of the primer may vary depending on various factors, for example, temperature and use of the primer. The primer may be 15 to 30 nucleotides in length. Short primer molecules generally require lower temperatures to form sufficiently stable hybrid complexes with the template.

The sequence of the primer does not need to have a sequence completely complementary to a partial sequence of the template, and it is sufficient if the complementarity within the range capable of hybridizing with the template and acting as a primer is inherent. Accordingly, the primer includes not only the polynucleotide itself but also a sequence that specifically hybridizes to the polynucleotide described above, which can act as a starting point in the polymerization reaction. For example, it may be a sequence completely complementary to the polynucleotide of SEQ ID NO: 1, or a sequence complementary to such a sequence that hybridizes to this sequence and can perform a primer action. The design of the primer can be readily carried out by those skilled in the art by reference to the sequence of the target nucleic acid to be amplified. For example, it can be designed using a commercially available program for primer design. Examples of commercially available programs for primer design include the PRIMER 3 program.

When the polynucleotide is used as a PCR primer, in addition to the polynucleotide, it may include a primer that specifically binds to its complementary strand.

"Probe" refers to a polynucleotide that specifically binds to a specific target sequence. The polynucleotide may be DNA or RNA. The polynucleotide may be in single stranded form. The polynucleotide may also be a polynucleotide which is not only composed of a natural nucleotide but also a natural nucleotide, an analogue of a natural nucleotide, a sugar, a base or a phosphate site of a natural nucleotide, which is capable of being hybridized to a complementary nucleotide by hydrogen bonding And nucleotides selected from the group consisting of combinations thereof. The polynucleotide includes PNA. Further, the polynucleotide may be attached with a detectable label (e.g., Cy3, Cy5 fluorescent substance), for example, in order to facilitate detection of the polynucleotide or the complex to which the polynucleotide is bound in the assay reaction, Terminal or 5 ' end.

The probe may be an entirely complementary sequence to the polynucleotide comprising the SNP site. In addition, the probe may have a substantially complementary sequence within a range that does not interfere with specific hybridization to the polynucleotide including the SNP site. In addition, the probe may have a modified nucleotide within a range that does not impair the specific hybridization to the polynucleotide including the SNP site. Examples of the probe include a perfect match probe consisting of a sequence completely complementary to the polynucleotide including a SNP site and a polynucleotide having a SNP site, And a probe having a completely complementary sequence.

The primers and probes can be used to amplify or confirm the presence of nucleotide sequences with specific alleles at the SNP site.

The polynucleotide may be a polynucleotide selected from the group consisting of the nucleotide sequences of SEQ ID NOS: 1-9 or a combination of complementary polynucleotides thereof.

Another embodiment provides a composition for diagnosing the risk of developing endogenous atopic dermatitis of an individual comprising the polynucleotide.

In the composition, the polynucleotide, the individual, and the endogenous atopic dermatitis are as described above. The composition may be a solid phase composition or a liquid phase composition. The liquid composition may comprise the polynucleotide in a suitable liquid medium. The liquid medium may be any of those used in the art as the liquid medium of the polynucleotide. In addition, the liquid medium may be appropriately selected depending on the intended use of the composition. For example, the liquid medium may be selected from the group consisting of water, PCR buffer, and hybridization buffer.

Another embodiment provides a microarray for diagnosing the risk of developing endogenous atopic dermatitis of an individual comprising the polynucleotide.

The above polynucleotide, individual, and endogenous atopic dermatitis are as described above. The term "microarray" means that the polynucleotide is immobilized at a high density in the divided region of the substrate surface. The microarray may be such that the regions are arranged on the substrate at a density of, for example, 400 / cm ² or more, 10 ³ / cm ² , or 10 ⁴ / cm ² .

Another embodiment provides a kit for diagnosing the risk of developing endogenous atopic dermatitis of an individual comprising the polynucleotide.

The above polynucleotide, individual and atopic dermatitis are as described above. The polynucleotides may be in the form of microarrays, arranged at high density in discrete regions on the substrate surface.

The kit may be a kit for specifically diagnosing the risk of developing atopic dermatitis of an individual through amplification of the polynucleotide and presence or absence of an amplification product. In this case, the kit may include the polynucleotide as a primer and the reagent necessary for amplification. The amplification reagent may comprise, for example, dNTPs, polymerases, and suitable buffers. For example, the primer may be one in which the nucleotide sequence corresponding to the SNP site forms the 3'-terminal nucleotide of the primer, the 3'-terminal nucleotide is complementary to the nucleotide at the SNP site (specific primer) (Non-specific primers). The non-specific primer may include a sequence that is not complementary to the 3 'terminal nucleotide as well as other sites. The kit may also include instructions for use. For example, in the case where the target sequence is amplified in the amplification reaction using the specific primer and the target sequence is not amplified in the amplification reaction using the non-specific primer, Including a description of determining the presence of a target sequence and determining the risk of developing an endogenous atopic dermatitis of an individual from the result.

The kit may be a kit for hybridizing the polynucleotide or a probe derived therefrom with a nucleic acid in a sample and diagnosing the risk of the endogenous atopic dermatitis of the individual from the hybridization result. In this case, the kit may include the probe and a reagent necessary for hybridization. Reagents necessary for hybridization include, for example, a hybridization buffer. The nucleic acid may not be amplified or amplified. Thus, the kit may further comprise reagents necessary for amplification of the nucleic acid. The nucleic acid may be labeled with a detectable label. The kit may comprise a perferct match probe that is completely complementary to the polynucleotide or a mismatch probe that is complementary at all sites except for the SNP site in the polynucleotide. The probes may be in the form of microarrays fixed to a plurality of discrete regions on the substrate. The kit may also include instructions for use. For example, when the target sequence is detected in the hybridization reaction using the completely complementary probe and the target sequence is not detected in the hybridization reaction using the mismatch probe, the sequence of the sequence associated with endogenous atopic dermatitis Including the description of determining the presence of atopic dermatitis, and determining the risk of developing an endogenous atopic dermatitis of the individual from the result.

Another embodiment provides a method for detecting nucleic acid, comprising: providing a separate nucleic acid sample; And

Determining the nucleotide at the SNP position of the polynucleotide as described above. ≪ Desc / Clms Page number 12 >

The method comprises providing a separate nucleic acid sample. Methods for separating nucleic acids from an individual are known in the art. For example, DNA can be isolated directly from tissues or cells by amplifying specific regions by nucleic acid amplification methods such as PCR. The separated nucleic acid sample includes not only the purely isolated nucleic acid but also a cell lysate containing a crude nucleic acid, for example, a nucleic acid. Such nucleic acid amplification methods include PCR, ligase chain reaction (LCR), transcription amplification, self-sustained sequence replication, and nucleic acid-based sequence amplification (NASBA). The separated nucleic acid may be DNA or RNA. The DNA may be a genomic DNA, a cDNA, or a recombinant DNA. The RNA may be mRNA.

The method also includes determining the nucleotide at the SNP position of the polynucleotide described above.

It is known to determine the nucleotide at the SNP position. For example, the nucleotide at the SNP position can be determined directly by a nucleotide sequencing method of known nucleic acids. The nucleotide sequence determination method may include a sanguine (or dideoxy) method or a curd-gilbert (chemical cleavage) method. In addition, the nucleotide at the SNP position can be determined by hybridizing a probe containing the sequence of the SNP position with the polynucleotide of interest, and analyzing the hybridization result. The degree of hybridization can be confirmed, for example, by marking a detectable label on the target nucleic acid, detecting the hybridized target nucleic acid, or confirming it by an electrical method or the like. In addition, a single base primer extension (SBE) method can be used.

In the above method, the step of determining a polynucleotide of the SNP region may include hybridizing the nucleic acid sample to a microarray on which the polynucleotide is immobilized; And detecting the hybridization result.

In addition, the method may include determining that the subject belongs to a high-risk group having an incidence of endogenous atopic dermatitis when a risk allele is present as a result of the determination of the nucleotide at the SNP site.

A "risk allele" is defined as a minor allele (a), a is a risk allele when the frequency of a in the disease group is greater than the frequency of a in the normal group, and vice versa A may be a risk allele. The more likely an individual has a risk allele, the greater the likelihood of endogenous atopic dermatitis. For example, in the case of SEQ ID NOS: 1, 7, 8 and 9, in the presence of a minority allele a, the subject is judged to belong to a high-risk group of endogenous atopic dermatitis, 3, 4, 5, and 6, in the presence of a minority allele a, the subject has a low probability of developing endogenous atopic dermatitis, or in the presence of major allele A, the possibility of endogenous atopic dermatitis It can be determined to belong to the risk group. Alleles A and a are shown in Table 1. For example, if the nucleotide at the determined SNP site is T for the SNP site of SEQ ID NO: 1, G for the SNP site of SEQ ID NO: 7, G for the SNP site of SEQ ID NO: 8, G for the SNP site of SEQ ID NO: G for the SNP region of SEQ ID NO: 3, C for the SNP region of SEQ ID NO: 4, T for the SNP region of SEQ ID NO: 5, and T for the SNP region of SEQ ID NO: The subject can be judged to belong to a high-risk group having a high incidence of endogenous atopic dermatitis. The nucleotide at the determined SNP site is G for the SNP site of SEQ ID NO: 1, T for the SNP site of SEQ ID NO: 7, A for the SNP site of SEQ ID NO: 8, A for the SNP site of SEQ ID NO: 2 for the SNP region, A for the SNP region of SEQ ID NO: 3, T for the SNP region of SEQ ID NO: 4, A for the SNP region of SEQ ID NO: 5, and C for the SNP region of SEQ ID NO: 6 , It can be determined that the subject belongs to a group having a low probability of developing endogenous atopic dermatitis.

In the method, the subject may be a human. The human being may be Korean. In the above method, the "altered risk" may be the relative risk of developing an endogenous atopic dermatitis. For example, the risk may be indicative of whether the probability of onset is increased or decreased relative to a normal group. The risk may also be indicative of whether the individual having a particular allele or genotype has an increased or decreased probability of developing endogenous atopic dermatitis as compared to an individual having another specific allele or genotype.

Table 1.

Table 1 shows the frequency of genotype and frequency of minor alleles in SNP sites in 212 patients with endogenous atopic dermatitis and 473 healthy individuals. The position in the sequence of the SNP shown in Table 1 is 101st.

In Table 1, the rs number is the SNP number registered in the NCBI (http://www.ncbi.nlm.nih.gov, national center for biotechnology information) GENBANK, and the sequence shown in the SEQ ID NO: Lt; RTI ID = 0.0 > SNP. ≪ / RTI > Those skilled in the art will readily be able to ascertain the location and sequence of the SNP with reference to the rs number. In Table 1, the sequence represented by the sequence number is a sequence including the SNP site as a part of the sequence represented by the NCBI Genbank registration number (rs number).

In Table 1, MA represents minor allele, and SNP represents the genotype or MAF at the SNP position. MAF_X (minor allele frequency) indicates the frequency of the minor allele X. Intrinsic atopic dermatitis and normal group represent genotype frequency or MAF in patients with endogenous atopic dermatitis and normal group, respectively.

In Table 1, "p-value" means a p-value which is the result of a test to determine whether the distribution of the genotype of the SNP is significant in a given model and in a patient group. "OR (95% CI)" represents the odds ratio, which is the probability of having the corresponding SNP among the patients in the normal group with respect to the probability of having the corresponding SNP based on the genotype. In the parentheses, % Means the lower and upper bounds of the 95% confidence interval taking into account the error range. If the odds ratio is greater than 1, the association of the minor allele of the SNP with endogenous atopic dermatitis increases the risk of developing an infection. If the odds ratio is less than 1, the association of the minor allele with endogenous atopic dermatitis Effect.

In other words, if the odds ratio is greater than 1, the minor allele of the SNP is a risk factor for endogenous atopic dermatitis. If the odds ratio is less than 1, the minor allele of the corresponding SNP acts as a protective factor against atopic dermatitis , Allergic factor is a risk factor for endogenous atopic dermatitis.

In Table 1, the significance model is a genetic model in which the minority alleles of SNPs are associated with the onset of endogenous atopic dermatitis. When dominant models have Aa or aa genotypes compared to those with AA genotype, endogenous atopy And the risk of developing dermatitis can be increased or decreased. That is, the risk of developing atopic dermatitis may be increased or decreased when one or two minority alleles are present, even when no minor allele is present.

Recessive is associated with an increase in the risk of endogenous atopic dermatitis when the aa genotype is compared to those with the AA and Aa genotypes, only when the two minor alleles are involved.

The codominant is characterized by an increase or decrease in the risk of endogenous atopic dermatitis as compared with the AA genotype when the genotype of the genotype Aa is compared to that of the genotype Aa.

Allele has an increased or decreased risk of developing endogenous atopic dermatitis by OR as compared to the A allele with an a allele.

The polynucleotides, compositions comprising them, microarrays and kits comprising them, according to one or more embodiments, can be used to rapidly and efficiently diagnose the risk of developing endogenous atopic dermatitis.

In addition, according to another embodiment, a method of identifying individuals with altered risk of developing endogenous atopic dermatitis can effectively identify whether an individual has an altered risk of developing endogenous atopic dermatitis.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example 1 Screening of SNP Markers Associated with Endogenous Atopic Dermatitis by Analysis of Allelic Frequency of SNP Locations in Patient and Normal Subjects

This example analyzed the allele frequency of SNP location in the genomes of endogenous atopic dermatitis patients and normal individuals and selected SNP markers associated with endogenous atopic dermatitis from the results.

(1) Classification of experimental subjects

In this example, a total of 1,118 patients, 212 patients with atopic dermatitis (female: 45.3%, mean age: 13.56 years) and 473 normal controls (female: 36.6%, mean age: 23.23 years) were collected.

Samples of patients with endogenous atopic dermatitis (AD) were collected from non-asthmatic atopic patients diagnosed at Samsung Medical Center (Seoul, Korea). Endogenous atopic dermatitis was diagnosed as endogenous atopic dermatitis when serum IgE levels were low in normal range and allergen specific IgE was not detected.

Normal controls were recruited from medical students and volunteers who did not have a history of skin injury due to atopic dermatitis.

(2) Analysis of genotypes through polarization detection

Genomic DNA was extracted from 5 ml of blood using a commercially available DNA separation kit (Gentra Genomic DNA purification kit, Minneapolis, MN, USA) according to the protocol provided by the manufacturer. Genotyping of SNP sites in the DNA was performed using the GenomeLab ^™ SNPstream ^® system (Ultra-high throughput; UHT system). The system utilizes tag sequence single nucleotide extension gene identification technology (Beckman Coulter, Fullerton, Calif., USA) in conjunction with multiplex PCR and SNPstream software accordingly.

The genotype analysis of the SNP site using the UHT system is summarized as follows. First, the sequences surrounding the 9 SNP regions shown in Table 1 were amplified from the genomic DNA by PCR. PCR was carried out simultaneously in each well of a 384-well plate using PCR primer pairs shown in Table 2 as primers, genomic DNA as a template, and Taq Gold polymerase as a polymerase. The thermocycles were repeated 34 times at 95 ° C for 30 seconds, at 55 ° C for 30 seconds and at 72 ° C for 1 minute.

Table 2.

Target sequence
(SEQ ID NO) Forward PCR
primer Reverse PCR
primer SNP-IT
primer One 10 11 12 2 13 14 15 3 16 17 18 4 19 20 21 5 22 23 24 6 25 26 27 7 28 29 30 8 31 32 33 9 34 35 36

Next, a single base primer extension reaction (also referred to as SNP-IT reaction) was performed using a primer (hereinafter also referred to as "SNP-IT primer") annealing immediately to the SNP. The sequences of the SNP-IT primers for each SNP are shown in Table 2. The SNP-IT primer contains a "tag" sequence at the 5 ' end of the primer. This tag sequence is annealed to a complementary probe nucleic acid (hereinafter referred to as a "tag probe") immobilized on a solid phase at the final stage of multiplex SNP-IT analysis. The SNP-IT reaction was first cleaved using exonuclease to the non-complementary strand for the SNP-IT primer. Next, a single base extension reaction was performed in each well of a 384 well plate in the presence of four differently-labeled differently-terminated dideoxynucleoside triphosphates, SNP-IT primer and DNA polymerase, each of which was different from each other. The heat cycle was repeated 45 times at 94 캜 for 20 seconds, and at 40 캜 for 11 seconds.

Next, the product of the single base extension reaction was added to each well of a SNPstream ^® Tag Array plate containing 384 wells and hybridized. Hybridization was carried out by incubation at 42 ° C for about 2 hours in a sucking incubator. There are sixteen divided regions on the surface of each well of the SNPstream ^® Tag Array plate, and a tag probe including a tag sequence is fixed in the divided region. Four of these have a tag sequence complementary to the control sequence and twelve have a complementary tag sequence to twelve single base extension products. Each SNP is identified by its well location.

Next, the hybridization results of the SNPstream using SNPstream UHT Array Imager ^{® TM?} The excitation light was applied to sixteen divided regions on the surface of each well of the Tag Array plate, and measurement was performed by measuring the radiation emitted therefrom. The measured light intensity is SNPstream ^? UHT computer and analyzed statistically.

(3) Statistical analysis

We used a chi-square test to determine whether individual variants follow the Hardy-Weinberg equilibrium (HWE) rule at each locus of the sample. Allelic effects (additional allelic effects) and genotype effects in individual SNPs were tested using a logistic regression model with sex and age as covariates. The p value of less than 0.05 was judged to be significant. Odds ratios were also measured by logistic regression analysis. Statistical analysis was performed using SAS 9.1 (SAS Institute Inc., NC, USA) and the R statistical language (http://www.r-project.org).

(4) Analysis results

The results of the analysis are shown in Table 1. As shown in Table 1, nine SNP markers associated with endogenous atopic dermatitis were identified. The nine SNP markers showed significantly different frequencies of alleles or genotypes between endogenous atopic dermatitis patients and normal individuals. SEQ ID NOS: 4 and 6 follow a dominant model, SEQ ID NO: 7 follows a recessive model, SEQ ID NOS: 1, 2, 3 and 9 are codominant, Followed by a model of significance, and SEQ ID NOS: 5 and 8 follow the model of allele oil.

In Table 1, the significance model is a genetic model in which the minority alleles of SNPs are associated with the onset of endogenous atopic dermatitis. When dominant models have Aa or aa genotypes compared to those with AA genotype, endogenous atopy And the risk of developing dermatitis can be increased or decreased. That is, the risk of developing atopic dermatitis may be increased or decreased when one or two minority alleles are present, even when no minor allele is present.

Recessive is associated with an increase in the risk of endogenous atopic dermatitis when the aa genotype is compared to those with the AA and Aa genotypes, only when the two minor alleles are involved.

The codominant is characterized by an increase or decrease in the risk of endogenous atopic dermatitis as compared with the AA genotype when the genotype of the genotype Aa is compared to that of the genotype Aa.

Allele is characterized by an increase or decrease in the risk of developing endogenous atopic dermatitis by OR as the a allele increases by one compared to the AA genotype.

In addition, the 9 SNP markers showed that the frequencies of minor alleles were not significantly different in all genetic models compared to normal subjects in the statistical analysis of the exogenous atopic dermatitis, another kind of atopic dermatitis, As a marker.

<110> DNA LINK, INC <120> Marker for diagnosing intrinsic atopic dermatitis and use thereof <130> PN085228 <160> 36 <170> Kopatentin 1.71 <210> 1 <211> 201 <212> DNA <213> Homo sapiens <220> <221> misc_feature <101> K represents g or t <400> 1 gtcaaactga acattgtcat aatacatgcc ggttgacacc aatcctctgc tcttccagcc 60 caggccttcc cttacgaggt ttaaaaacat gctgaagggt kgaagctacc aagggtcttc 120 tggtgttttt ctcccttcct ctgaagtgca taattattct gtttactgga agattttcaa 180 cacaaaggaa attaacttct c 201 <210> 2 <211> 201 <212> DNA <213> homo sapiens <220> <221> misc_feature <101> <223> r represents a or g <400> 2 taaatttcaa gccttgtcct ttcgtaaata gtcattgcaa agacaaagaa ggtgtattta 60 acaaacttga attgtttcca ggtcacccat ttgctgtagc rgtgtttgag gattatgtgt 120 ggttctcaga ttgggctatg ccatcagtaa tgagagtaaa caagaggact ggcaaagata 180 gagtacgtct ccaaggcagc a 201 <210> 3 <211> 201 <212> DNA <213> homo sapiens <220> <221> misc_feature <101> <223> r represents a or g <400> 3 tactaaaatt taagtacttg tgtgggagag aagaaatatg aagataacta ggaacataac 60 tttagatgtt taaggcagtc ttttctgcag aactcaaaat rctttccaga cagtgtttca 120 ctctttccaa ggttctgtga ggtagagggt gtgtgtgtgt ttgtgtgtgt gtgtgtgtgt 180 gtgtgtgtgt gtgtgtgtat g 201 <210> 4 <211> 201 <212> DNA <213> homo sapiens <220> <221> misc_feature <101> <223> y represents c or t <400> 4 ggggtgtggg atagaaggca ggtgtccttg aatcatgacc tagcctcagc atccctctaa 60 attagttgca tgtaacccaa tctcccccac ctaggcttca ytgtgtttca atctttttga 120 ataaagcggg ccatctttct ttctgtgtgt gaccttggag ttacacataa gcagaataga 180 tgtgcatcta ttctccatct a 201 <210> 5 <211> 201 <212> DNA <213> homo sapiens <220> <221> misc_feature <101> <223> w represents a or t <400> 5 tcctgggcta aagagatcct cccatttcaa cttccagagc agctgggaca acaggcgctt 60 gccaccacac ctggctaatt ttcttatttt aatttaattt wattttattt tttgggacag 120 agtggagtct caaaaaccaa gctggagtgc agtggtgcga tctcgactca ctgcaatctc 180 tgcctcccgg gttcaagcga t 201 <210> 6 <211> 201 <212> DNA <213> homo sapiens <220> <221> misc_feature <101> <223> y represents c or t <400> 6 tggcttcagg gtaagaactg attaagaagg aaaaagggaa ggccctgcag aggaggacta 60 tgagcaaggc tggtaggatg agcgtaccat ggactatcca yaaactcatt gggatattaa 120 ggtcaaaagt agccatgtgg catctccatt ggacagcagc aagttccaac tagaaaaatc 180 tcgggaaata cttcatgatt a 201 <210> 7 <211> 201 <212> DNA <213> homo sapiens <220> <221> misc_feature <101> K represents g or t <400> 7 caaagcagag aaaacacacc ccctcggtgc tttcctttac ttatgggaga cgtggaatgc 60 tttgcactat tccagcaaca agattcagga ggaacatcga kaaattggag ggaaccacca 120 gggctgctgt gtggaaaggc tgaaggccag agaagcagag gcacaggagg gcagaagagg 180 aaggcaaagg aaggtctcag c 201 <210> 8 <211> 201 <212> DNA <213> homo sapiens <220> <221> misc_feature <101> <223> r represents a or g <400> 8 gtggttgggc ctgacggatt taaaccagct aaaaggtcag agccaaaata agtgctattc 60 cctaatggaa acctccctaa atttctaaaa cactgtgtac rtgcccttca aaatttgata 120 aagattcact aacaaaacag tttttcttaa ttgaattttc catgtctact acactttctg 180 aaaaataaaa ctgtttgtgt g 201 <210> 9 <211> 201 <212> DNA <213> homo sapiens <220> <221> misc_feature <101> <223> r represents a or g <400> 9 accaccgtcc cctcagccgc tcaagccaag caccctgggg ttctccttgc ttcctccttc 60 ccctcactct tcaaagccaa tccatccaga agtcctgtcc rttctaactc caggacacat 120 ccgtgcatct ccattcctgg ccctcctgcg gagccagctg tgtccttgcc ggctctgtga 180 catcctcccg cggactgctg t 201 <210> 10 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> forward primer <400> 10 cagcccaggc cttccctta 19 <210> 11 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> reverse primer <400> 11 ttcagaggaa gggagaaaaa c 21 <210> 12 <211> 45 <212> DNA <213> Artificial Sequence <220> <223> SNP-IT primer <400> 12 agagcgagtg acgcatacta gaggtttaaa aacatgctga agggt 45 <210> 13 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> forward primer <400> 13 aaggtgtatt taacaaactt gaattg 26 <210> 14 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> reverse primer <400> 14 atggcatagc ccaatctg 18 <210> 15 <211> 45 <212> DNA <213> Artificial Sequence <220> <223> SNP-IT primer <400> 15 gcggtaggtt cccgacatat ttccaggtca cccatttgct gtagc 45 <210> 16 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> forward primer <400> 16 accctctacc tcacagaacc tt 22 <210> 17 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> reverse primer <400> 17 ttgtgtggga gagaagaaat atg 23 <210> 18 <211> 45 <212> DNA <213> Artificial Sequence <220> <223> SNP-IT primer <400> 18 gacctgggtg tcgataccta aaagagtgaa acactrtctg gaaag 45 <210> 19 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> forward primer <400> 19 agcatccctc taaattagtt gc 22 <210> 20 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> reverse primer <400> 20 tagatgcaca tctattctgc ttatg 25 <210> 21 <211> 45 <212> DNA <213> Artificial Sequence <220> <223> SNP-IT primer <400> 21 gcggtaggtt cccgacatat cccaatctcc cccacctagg cttca 45 <210> 22 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> forward primer <400> 22 ygcttgccac cacacctg 18 <210> 23 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> reverse primer <400> 23 tcyagcttgg tttttgagac tc 22 <210> 24 <211> 45 <212> DNA <213> Artificial Sequence <220> <223> SNP-IT primer <400> 24 gacctgggtg tcgataccta taattttctt attttaattt aattt 45 <210> 25 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> forward primer <400> 25 agaggaggac tatgagcaag g 21 <210> 26 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> reverse primer <400> 26 aatggagatg ccacatgg 18 <210> 27 <211> 45 <212> DNA <213> Artificial Sequence <220> <223> SNP-IT primer <400> 27 gcggtaggtt cccgacatat ttgaccttaa tatcccaatg agttt 45 <210> 28 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> forward primer <400> 28 ccttcagcct ttccacac 18 <210> 29 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> reverse primer <400> 29 aaagcagaga aaacacaccc 20 <210> 30 <211> 45 <212> DNA <213> Artificial Sequence <220> <223> SNP-IT primer <400> 30 agagcgagtg acgcatacta cagccctggt ggttccctcc aattt 45 <210> 31 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> forward primer <400> 31 aaaataagtg ctattcccta atgga 25 <210> 32 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> reverse primer <400> 32 tagtagacat ggaaaattca attaagaa 28 <210> 33 <211> 45 <212> DNA <213> Artificial Sequence <220> <223> SNP-IT primer <400> 33 agggtctcta cgctgacgat cctaaatttc taaaacactg tgtac 45 <210> 34 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> forward primer <400> 34 ttcctccttc ccctcact 18 <210> 35 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> reverse primer <400> 35 agggccagga atggagat 18 <210> 36 <211> 45 <212> DNA <213> Artificial Sequence <220> <223> SNP-IT primer <400> 36 agatagagtc gatgccagct gccaatccat ccagaagtcc tgtcc 45

Claims (10)

1. A polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1 or a complementary polynucleotide thereof, wherein the polynucleotide comprises the SNP position of the polynucleotide and comprises 10 or more consecutive nucleotides selected from the polynucleotide, A composition for diagnosing the risk of developing endogenous atopic dermatitis of an individual comprising a polynucleotide for diagnosing the risk of developing an endogenous atopic dermatitis of an individual that is the 101st nucleotide of SEQ ID NO: 1. 2. The composition of claim 1, wherein the polynucleotide is a polynucleotide having a length of from 10 to 100 nucleotides. 2. The composition of claim 1, wherein the polynucleotide is a polynucleotide that is a primer or a probe. delete 1. A polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1 or a complementary polynucleotide thereof, wherein the polynucleotide comprises the SNP position of the polynucleotide and comprises 10 or more consecutive nucleotides selected from the polynucleotide, A microarray for diagnosing the risk of developing endogenous atopic dermatitis of an individual comprising a polynucleotide for diagnosing the risk of developing an endogenous atopic dermatitis of an individual that is the 101st nucleotide of SEQ ID NO: 1. 1. A polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1 or a complementary polynucleotide thereof, wherein the polynucleotide comprises the SNP position of the polynucleotide and comprises 10 or more consecutive nucleotides selected from the polynucleotide, A kit for diagnosing the risk of developing an endogenous atopic dermatitis of an individual comprising a polynucleotide for diagnosing the risk of developing endogenous atopic dermatitis of an individual that is the 101st nucleotide of SEQ ID NO: 1. Providing an isolated nucleic acid sample; And 1. A polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1 or a complementary polynucleotide thereof, wherein the polynucleotide comprises the SNP position of the polynucleotide and comprises 10 or more consecutive nucleotides selected from the polynucleotide, Determining the nucleotide at the SNP position of the polynucleotide to diagnose the risk of developing an endogenous atopic dermatitis of an individual that is the 101st nucleotide of SEQ ID NO: 1. 8. The method according to claim 7, wherein, when a risk allele is present as a result of the determination of the nucleotide at the SNP site, the step of determining that the subject belongs to a risk group having a high risk of developing endogenous atopic dermatitis. 8. The method of claim 7, comprising determining if the nucleotide at the determined SNP site is a T nucleotide for the SNP site of SEQ ID NO: 1, the subject is determined to belong to a high risk group of endogenous atopic dermatitis. 8. The method of claim 7, comprising determining if the nucleotide at the determined SNP site is a G nucleotide for the SNP site of SEQ ID NO: 1, the subject is determined to belong to a group having a low probability of developing endogenous atopic dermatitis.