KR20230025094A - SNP for drug hypersensitivity and diagnosis method using the same - Google Patents

Abstract

The present invention relates to an SNP for the diagnosis of drug hypersensitivity reactions and a diagnosis method using the same. As a result of analyzing a protein coding region of genomic DNA of patients with drug hypersensitivity reactions, inventors of the present invention have found that a specific SNP containing rs35372932 is closely related to the onset of drug hypersensitivity reactions. Therefore, the SNP can be used to develop a genetic diagnostic reagent for the diagnosis or prediction of drug hypersensitivity reactions.

Description

SNP for drug hypersensitivity diagnosis and diagnosis method using the same {SNP for drug hypersensitivity and diagnosis method using the same}

The present invention relates to a SNP for diagnosing a drug hypersensitivity reaction and a diagnostic method using the same.

Medically speaking, drug allergy is a reaction mediated by an immunological mechanism in a patient sensitized to a drug, which is different from saying that the general public or a patient has drug side effects or drug allergy. In general, adverse drug reactions or drug allergies often mean adverse drug reactions. Adverse drug reactions include most of the adverse reactions caused by drugs. It was defined as an unintended harmful reaction to the human body in addition to the therapeutic effect when a normal dose of a drug is administered through an appropriate route of administration. Adverse drug reactions can be largely divided into predictable A-type reactions that occur in relation to the dose of the drug and unpredictable B-type reactions that occur regardless of the dose of the drug. Predictable A-type adverse drug reactions are mainly caused by the unique pharmacological action of the drug, about 80% of the total adverse drug reactions, so they include drug addiction, side effects, indirect effects, and drug interactions in a dose-dependent manner. Unpredictable type B adverse drug reactions account for the remaining 15-20%, and include drug-induced hypersensitivity, idiosyncrasy, intolerance, and pseudoallergic reactions. .

The World Allergy Organization (WAO) recommends classifying drug allergy into immediate (within 1 hour) and delayed (after 1 hour), depending on the time taken for symptoms to appear after drug exposure. According to the Gell & Coombs classification, hypersensitivity reactions not only to drugs but also to other environmental factors are divided into four types. Of these, those related to drug allergy are mainly IgE-related type I and T-cell involved type IV.

Type I is an immediate type, which is a humoral immune response mediated by IgE, mast cells, and basophils, and causes anaphylaxis, angioederma, bronchospasm, urticaria, and hives. belong to Typical drugs that show this type of reaction are β-lactam antibiotics (penicillins and cephalosporins), muscle-nervous system blocking drugs quinolones, platinum compounds such as carboplatin and oxaliplatinn, and cetuximab. and monoclonal antibodies such as rituximab.

Types II to IV are the delayed type, and type II refers to a humoral cytotoxic reaction mediated by IgM or IgG antibodies and complement, and includes hemolytic anemia, thrombocytopenia, and neutropenia. ) and so on. Typical drugs include β-lactam antibiotics (penicillins and cephalosporins), NSAIDs (Non-steroidal Antiinflammtory Drugs), quinine/quinidine, vancomycin, carbamazepine, propylthiouracil, and flecainide. (flecainide), etc.

As such, drug hypersensitivity reactions are the most serious form of allergic reactions and are regarded as a medical emergency, so it is very important to predict the occurrence of drug hypersensitivity reactions in advance.

Accordingly, the present inventors performed GWAS analysis to predict and diagnose drug hypersensitivity in advance, and identified SNPs having a significant correlation with drug hypersensitivity, thereby completing the present invention.

Kim Soo-jin : Drug Allergy, Journal of Hospital Pharmacists (2011), Vol. 28, No. 4

이에, 본 발명의 목적은 dbSNP 데이터베이스 rs35372932, rs79736818, rs3129871, rs9557291, rs11724428, rs11123552, rs62124613, rs77389536, rs206887084, rs9557291, rs372025, rs8058114, rs143395859, rs57833801, rs139830888, rs9384423, rs6097064, rs2175562, rs3924164, rs11206477, rs62225078 , rs73202933, rs72622187, rs139141104, rs9994092, rs1762852884, rs2335545, rs9688895, rs144130219, rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, 및 rs28717999로 이루어진 군으로부터 선택된 하나 이상의 SNP의 검출 제제를 포함 To provide a composition for diagnosing or predicting the onset of a drug hypersensitivity reaction.

Another object of the present invention is to provide a kit for diagnosing drug hypersensitivity including the composition.

본 발명의 또 다른 목적은 피검체로부터 수득한 시료 중의 SNP 부위의 염기서열에 dbSNP database rs35372932, rs79736818, rs3129871, rs9557291, rs11724428, rs11123552, rs62124613, rs77389536, rs206887084, rs9557291, rs372025, rs8058114, rs143395859, rs57833801, rs139830888, rs9384423, rs6097064, rs2175562, rs3924164, rs11206477, rs62225078, rs73202933, rs72622187, rs139141104, rs9994092, rs1762852884, rs2335545, rs9688895, rs144130219, rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, 및 rs28717999 When one or more single nucleotide polymorphisms selected from the group consisting of are present, it is to provide a method for providing information necessary for diagnosing a drug hypersensitivity reaction comprising determining that the subject has a drug hypersensitivity reaction.

Another object of the present invention is to dbSNP database rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs144157017, rs144157017, rs13250548, rs144157017, rs144157017, rs59594955, and rs295. It is an object of the present invention to provide a method for providing information necessary for predicting the onset of a drug hypersensitivity reaction, including determining that a subject has a high risk of developing a drug hypersensitivity reaction when a polymorphism exists.

본 발명의 또 다른 목적은 약물 과민반응 환자로부터 수득한 시료 중의 SNP 부위의 염기서열에 dbSNP database rs35372932, rs79736818, rs3129871, rs9557291, rs11724428, rs11123552, rs62124613, rs77389536, rs206887084, rs9557291, rs372025, rs8058114, rs143395859, rs57833801, rs139830888, rs9384423, rs6097064, rs2175562, rs3924164, rs11206477, rs62225078, rs73202933, rs72622187, rs139141104, rs9994092, rs1762852884, rs2335545, rs9688895, rs144130219, rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, And if at least one single nucleotide polymorphism selected from the group consisting of rs28717999 is present, determining that the subject has a delayed drug hypersensitivity reaction To provide a method for providing information necessary for diagnosis of a delayed drug hypersensitivity reaction will be.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

상기 과제를 해결하기 위하여, 본 발명은 dbSNP 데이터베이스 rs35372932, rs79736818, rs3129871, rs9557291, rs11724428, rs11123552, rs62124613, rs77389536, rs206887084, rs9557291, rs372025, rs8058114, rs143395859, rs57833801, rs139830888, rs9384423, rs6097064, rs2175562, rs3924164, rs11206477, rs62225078, rs73202933, rs72622187, rs139141104, rs9994092, rs1762852884, rs2335545, rs9688895, rs144130219, rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, 및 rs28717999로 이루어진 군으로부터 선택된 하나 이상의 SNP의 검출 Provided is a composition for diagnosing or predicting the onset of a drug hypersensitivity reaction comprising an agent.

In one embodiment of the present invention, the drug hypersensitivity reaction may be an immediate drug hypersensitivity reaction or a delayed drug hypersensitivity reaction, but is not limited thereto.

In another embodiment of the present invention, the SNP is selected from the group consisting of i) rs35372932, rs79736818, rs3129871, rs9557291, rs11724428, rs11123552, rs62124613, rs77389536, and rs206887084 If the composition is at least one hypersensitivity diagnosis, or for predicting onset,

If the SNP is one or more selected from the group consisting of ii) rs9557291, rs372025, rs8058114, rs143395859, rs57833801, rs139830888, rs9384423, rs6097064, rs2175562, and rs3924164 for diagnosis or predicting normal onset drug, the composition is is,

If the SNP is one selected from the group consisting of: iii) rs11206477, rs62225078, rs73202933, rs72622187, rs139141104, rs9994092, rs1762852884, rs2335545, rs9688895, and rs144130219 for predicting a hypersensitivity-onset reaction or abnormal drug, the composition for normal diagnosis or abnormality is,

If the SNP is at least one selected from the group consisting of iv) rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, and rs28717999, the composition for distinguishing immediate drug hypersensitivity reaction from immediate type drug hypersensitivity reaction, It may be, but is not limited thereto.

In another embodiment of the present invention, the SNP may be rs35372932 or rs139141104, but is not limited thereto.

In another embodiment of the present invention, the drug hypersensitivity reaction may have occurred in Koreans, but is not limited thereto.

In another embodiment of the present invention, the detection agent may be a probe capable of detecting the SNP, but is not limited thereto.

In another embodiment of the present invention, the detection agent may be a primer capable of detecting the SNP, but is not limited thereto.

In addition, the present invention provides a kit for diagnosing drug hypersensitivity including the composition.

In one embodiment of the present invention, the kit may be a RT-PCR kit or a microarray chip kit, but is not limited thereto.

또한, 본 발명은 피검체로부터 수득한 시료 중의 SNP 부위의 염기서열에 dbSNP database rs35372932, rs79736818, rs3129871, rs9557291, rs11724428, rs11123552, rs62124613, rs77389536, rs206887084, rs9557291, rs372025, rs8058114, rs143395859, rs57833801, rs139830888, rs9384423, rs6097064, rs2175562, rs3924164, rs11206477, rs62225078, rs73202933, rs72622187, rs139141104, rs9994092, rs1762852884, rs2335545, rs9688895, rs144130219, rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, 및 rs28717999으로 이루어진 Provided is a method for providing information necessary for diagnosing a drug hypersensitivity reaction comprising determining that a subject has a drug hypersensitivity reaction when one or more single nucleotide polymorphisms selected from the group exist.

또한, 본 발명은 피검체로부터 수득한 시료 중의 SNP 부위의 염기서열에 dbSNP database rs35372932, rs79736818, rs3129871, rs9557291, rs11724428, rs11123552, rs62124613, rs77389536, rs206887084, rs9557291, rs372025, rs8058114, rs143395859, rs57833801, rs139830888, rs9384423, rs6097064, rs2175562, rs3924164, rs11206477, rs62225078, rs73202933, rs72622187, rs139141104, rs9994092, rs1762852884, rs2335545, rs9688895, rs144130219, rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, 및 rs28717999으로 이루어진 Provided is a method for providing information necessary for predicting the onset of a drug hypersensitivity reaction, comprising determining that the subject has a high risk of drug hypersensitivity reaction when one or more single nucleotide polymorphisms selected from the group exist.

In addition, the present invention dbSNP database rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, and rs2985 selected from a single group consisting of rs1276175, rs144157017, rs59594955, and rs2987 Provided is a method for providing information necessary for diagnosis of a delayed drug hypersensitivity reaction, comprising determining that the subject has a delayed drug hypersensitivity reaction when the polymorphism exists.

In one embodiment of the present invention, the method includes sequencing, exome sequencing, microarray hybridization, allele specific PCR, and dynamic allele hybridization techniques ( It may be performed by one or more methods selected from the group consisting of dynamic allele-specific hybridization), PCR extension analysis, and Taqman technique, but is not limited thereto.

또한, 본 발명은 dbSNP database rs35372932, rs79736818, rs3129871, rs9557291, rs11724428, rs11123552, rs62124613, rs77389536, rs206887084, rs9557291, rs372025, rs8058114, rs143395859, rs57833801, rs139830888, rs9384423, rs6097064, rs2175562, rs3924164, rs11206477, rs62225078, rs73202933 , rs72622187, rs139141104, rs9994092, rs1762852884, rs2335545, rs9688895, rs144130219, rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, 및 rs28717999으로 이루어진 군으로부터 선택된 하나 이상의 SNP의 검출 제제를 포함하는 조성물 It provides diagnostic use of drug hypersensitivity reactions.

또한, 본 발명은 dbSNP database rs35372932, rs79736818, rs3129871, rs9557291, rs11724428, rs11123552, rs62124613, rs77389536, rs206887084, rs9557291, rs372025, rs8058114, rs143395859, rs57833801, rs139830888, rs9384423, rs6097064, rs2175562, rs3924164, rs11206477, rs62225078, rs73202933 , rs72622187, rs139141104, rs9994092, rs1762852884, rs2335545, rs9688895, rs144130219, rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, 및 rs28717999으로 이루어진 군으로부터 선택된 하나 이상의 SNP의 검출 제제를 포함하는 조성물 It provides use for predicting the onset of drug hypersensitivity reactions.

본 발명자들은 약물 과민반응 환자들의 유전체 DNA의 단백질 코딩 부위를 분석한 결과, 단일염기다형성(SNP) 부위인 dbSNP database rs35372932, rs79736818, rs3129871, rs9557291, rs11724428, rs11123552, rs62124613, rs77389536, rs206887084, rs9557291, rs372025 , rs8058114, rs143395859, rs57833801, rs139830888, rs9384423, rs6097064, rs2175562, rs3924164, rs11206477, rs62225078, rs73202933, rs72622187, rs139141104, rs9994092, rs1762852884, rs2335545, rs9688895, rs144130219, rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175 , rs144157017, rs59594955, and rs28717999 were found to be closely related to the onset of drug hypersensitivity reactions, and it can be used to develop a genetic diagnostic reagent for diagnosing or predicting the onset of drug hypersensitivity reactions.

Figures 1a and 1b summarize the quality control (QC) process flow according to an embodiment of the present invention, Figure 1a is a quality control process flow in discovery analysis, Figure 1b shows a quality control process flow in replication analysis .
Figure 2 shows the 1000G dataset as a multidimensional scaling (MDS) plot of discovery data, with blue, yellow, and gray representing the control, case (DHR, immediate + delayed), and 1000G populations, respectively.
Figures 3a to 3c show the quartile (QQ) plots of the assay data, Figure 3a shows the control versus DHR, Figure 3b control versus immediate response, Figure 3c shows the control versus delayed response analysis results.
Figure 4 shows a Manhattan plot of control versus DHR (immediate + delayed) according to discovery analysis, where the red line represents the Bonferroni-adjusted significance level.
5A and 5B show area plots, FIG. 5A is the logistic regression analysis results for control versus DHR, and FIG. 5B shows a generalized multinomial logit model for control versus delayed response.
Figure 6 shows a Miami plot according to the discovery analysis, the upper part shows the comparison result of the immediate response and the control group, and the lower part shows the comparison result of the delayed response and the control group. The dotted line represents the Bonferroni-adjusted significance level.
Figure 7 shows the ratio of phenotypic variation explained by genotypic SNPs according to different prevalence, and the ratio of phenotypic variation explained by genotypic SNP, hSNP ² was calculated by GCTA.

본 발명은 dbSNP database rs35372932, rs79736818, rs3129871, rs9557291, rs11724428, rs11123552, rs62124613, rs77389536, rs206887084, rs9557291, rs372025, rs8058114, rs143395859, rs57833801, rs139830888, rs9384423, rs6097064, rs2175562, rs3924164, rs11206477, rs62225078, rs73202933, rs72622187 , rs139141104, rs9994092, rs1762852884, rs2335545, rs9688895, rs144130219, rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, 및 rs28717999으로 이루어진 군으로부터 선택된 하나 이상의 SNP의 검출 제제를 포함하는 약물 과민반응 It relates to a composition for diagnosis or onset prediction of

Hereinafter, the present invention will be described in detail.

As used herein, “polynucleotide” or “nucleic acid” refers to deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) in single or double-stranded form. Unless otherwise limited, known analogs of natural nucleotides that hybridize to nucleic acids in a manner similar to naturally occurring nucleotides are also included. In general, DNA is composed of four bases: adenine (A), guanine (G), cytosine (C), and thymine (T), and RNA uses uracil (U) instead of thymine. has In the nucleic acid double strand, A forms a hydrogen bond with T or U, and C with G base, and the relationship between these bases is called 'complementary'.

In this specification, when expressing a mutation of a gene, 'c.' means a mutation in a protein coding region, and '(base position) (base one letter) (symbol >) (base one letter)' is at the corresponding base position. It means that the preceding marked base is substituted with the following marked base.

As used herein, 'polymorphism' means the occurrence of two or more alternative sequences or alleles (or alleles) within a genetically determined population, and 'single nucleotide polymorphism (SNP)' refers to one Refers to the polymorphism of bases. Specifically, a single base (A, T, C, or G) in the genome refers to the diversity of DNA sequences occurring between members of a species or between paired chromosomes of an individual. For example, when three DNA fragments from different individuals contain differences in a single base, such as AAGT[A/A]AG, AAGT[A/G]AG, and AAGT[G/G]AG , called two alleles (A or G), and generally almost all SNPs have two alleles. In addition, when an SNP is genetically closely related to a specific disease, the SNP is a mutation in one base at a specific position compared to an identified normal or wild-type (WT) individual or allele. it also means

On the other hand, 'mRNA (messenger RNA or messenger RNA)' is RNA that serves as a blueprint for polypeptide synthesis (protein translation) by transferring the genetic information of the nucleotide sequence of a specific gene to ribosome during protein synthesis. Using a gene as a template, single-stranded mRNA is synthesized through a transcription process.

As used herein, 'protein' is used interchangeably with 'polypeptide' or 'peptide', and refers to a polymer of amino acid residues, eg as commonly found in proteins in nature.

As used herein, the first letter (three letter) of an amino acid refers to the following amino acids according to standard abbreviation conventions in the field of biochemistry: A (Ala): alanine; C(Cys): cysteine; D(Asp): aspartic acid; E (Glu): glutamic acid; F(Phe): phenylalanine; G(Gly): glycine; H(His): histidine; I(IIe): isoleucine; K(Lys): lysine; L (Leu): Leucine; M (Met): methionine; N(Asn): Asparagine; O(Ply): pyrrolysine; P(Pro): proline; Q(Gln): glutamine; R(Arg): arginine; S (Ser): serine; T(Thr): threonine; U(Sec):selenocysteine; V(Val): valine; W(Trp): tryptophan; Y (Tyr): Tyrosine.

'(amino acid letter) (amino acid position) (amino acid letter)' used herein means that the preceding amino acid at the corresponding amino acid position of the wild-type protein is replaced with the succeeding amino acid.

As used herein, the term “complementary” refers to a targeting moiety in a nucleic acid molecule that is capable of binding to a target (e.g., a SNP of the present invention) under predetermined hybridization or annealing conditions, specifically physiological conditions (intracellular). Sufficiently complementary to hybridize selectively, possibly having one or more mismatched base sequences, encompassing both substantially complementary and perfectly complementary sequences. , and more specifically means completely complementary.

본 명세서에서 상기 조성물은 dbSNP database rs35372932, rs79736818, rs3129871, rs9557291, rs11724428, rs11123552, rs62124613, rs77389536, rs206887084, rs9557291, rs372025, rs8058114, rs143395859, rs57833801, rs139830888, rs9384423, rs6097064, rs2175562, rs3924164, rs11206477, rs62225078, rs73202933, rs72622187, rs139141104, rs9994092, rs1762852884, rs2335545, rs9688895, rs144130219, rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, 및 rs28717999으로 이루어진 군으로부터 선택된 하나 이상의 SNP를 진단 마커로 검출한다 .

In the present specification, SNP rs35372932 is a substitution of 32597208th base on chromosome 6 from C to T.

In the present specification, SNP rs79736818 is a substitution of the 148357581st base on chromosome 5 from A to G.

In the present specification, SNP rs3129871 is a substitution of A at the 32438565th base on chromosome 6 from C.

In this specification, SNP rs9557291 is 99761210th base on chromosome 13 is substituted from G to A.

In the present specification, SNP rs11724428 is a substitution of A from G at base 112598451 on chromosome 4.

In the present specification, SNP rs11123552 is a substitution of G from C at the 120454245th base on chromosome 2.

In the present specification, SNP rs62124613 is a substitution of the 20386466th base on chromosome 2 from C to T.

In the present specification, SNP rs77389536 is a substitution of base 102408368 from T to C on chromosome 10.

In the present specification, SNP rs206887084 is a substitution of A from G at the 94620980th base on chromosome 12.

In this specification, SNP rs9557291 is 99761210th base on chromosome 13 is substituted from G to A.

In the present specification, SNP rs372025 is a substitution of the 107077214th base on chromosome 4 from T to C.

In the present specification, SNP rs8058114 is a substitution of A from G at the 12603772nd base on chromosome 16.

In the present specification, SNP rs143395859 is a substitution of A from G at the 27293404th base on chromosome 7.

In the present specification, SNP rs57833801 is a substitution of A from G at the 121777405th base on chromosome 2.

In the present specification, SNP rs139830888 is a substitution of A from G at the 147872742nd base on chromosome 7.

In the present specification, SNP rs9384423 is a substitution of base 156089071 from A to G on chromosome 6.

In the present specification, SNP rs6097064 is a substitution of base 37952348 from A to C on chromosome 20.

In the present specification, SNP rs2175562 is a substitution of the 140398639th base on chromosome 4 from A to G.

In the present specification, SNP rs3924164 is a substitution of the 76737927th base on chromosome 18 from C to T.

In this specification, SNP rs11206477 is 54941621st base on chromosome 1 is substituted from C to A.

In the present specification, SNP rs62225078 is 47740975 base on chromosome 22 is substituted from T to A.

In the present specification, SNP rs73202933 is a substitution of the 89819306th base on chromosome 12 from T to C.

In the present specification, SNP rs72622187 is a substitution of A from G at the 6078051st base on chromosome 3.

In the present specification, SNP rs139141104 is a substitution of base 31021244 from A to G on chromosome 6.

In the present specification, SNP rs9994092 is 65570396 base on chromosome 4 is substituted from C to A.

In the present specification, SNP rs1762852884 is a substitution of the 30994936th base on chromosome 6 from A to C.

In the present specification, SNP rs2335545 is a substitution of the 2916111th base on chromosome 16 from C to T.

In this specification, SNP rs9688895 is 38465523 base on chromosome 6 is substituted from C to A.

In the present specification, SNP rs144130219 is a substitution of the 98725782nd base on chromosome 13 from T to C.

In the present specification, SNP rs78831487 is a substitution of the 6537140th base on chromosome 19 from T to C.

In the present specification, SNP rs456449 is a substitution of A from G at the 38851003rd base on chromosome 21.

In the present specification, SNP rs13250548 is a substitution of base 35650882 from A to G on chromosome 8.

In the present specification, SNP rs77068773 is a substitution of the 23204196th base on chromosome 21 from A to T.

In the present specification, SNP rs3025035 is a substitution of the 43783622nd base on chromosome 6 from C to T.

In this specification, SNP rs2088142 is a substitution of A from G at the 33691285th base on chromosome 15.

In the present specification, SNP rs1276175 is a substitution of A from G at the 30945691st base on chromosome 5.

In the present specification, SNP rs144157017 is a substitution of base 50740465 from A to G on chromosome 6.

In the present specification, SNP rs59594955 is a substitution of base 38443346 from A to C on chromosome 6.

In the present specification, SNP rs28717999 is 65571223 base on chromosome 4 is substituted from A to G.

The above-mentioned SNPs may be located on the nucleotide sequences of SEQ ID NO: 1 to SEQ ID NO: 38, respectively. In the base sequences of SEQ ID NO: 1 to SEQ ID NO: 38 mentioned above, the base corresponding to the SNP position according to the present invention is represented by r, and based on the SNP position, the sequence 10 nt in the 5' direction, the sequence in the 3' direction It shows the sequence 10 nt. For example, in the nucleotide sequence of SEQ ID NO: 1 (rs_id, rs35372932), the SNP position is denoted by r and 10 nt long nucleotide sequences are displayed on both sides based on this, and a sequence having a total length of 21 nt is displayed. In addition, when the base indicated by r is identified as, for example, the majority allele T or the minor allele C, it is indicated as r = t or c in the identification number <223> section of the sequence listing. The nucleotide sequences of SEQ ID NO: 1 to SEQ ID NO: 38 are used for the purpose of specifying the position of a SNP in an individual, particularly a human, and therefore, the length of the nucleotide sequences of SEQ ID NO: 1 to SEQ ID NO: 38 or the indicated range The present invention is not limited by

In this specification, the term “detection” means both measuring and confirming the presence (expression) of a target substance or measuring and confirming a change in the presence (expression level) of a target substance. In the same context, measuring the expression level of the SNP in the present invention means measuring whether or not it is expressed (ie, measuring whether or not it is expressed) or measuring the level of qualitative or quantitative change of the SNP. The measurement can be performed without limitation including both qualitative methods (analysis) and quantitative methods. Types of qualitative and quantitative methods for determining the presence of SNPs are well known in the art, and the experimental methods described herein are included. A specific SNP level comparison method for each method is well known in the art.

In the present specification, “drug hypersensitivity” is also referred to as drug allergy, and refers to a reaction mediated by an immunological mechanism in a patient sensitized to a drug. Drug allergy is a case where an adverse reaction occurs by an immune mechanism regardless of the drug's original pharmacological action, and accounts for about 6-10% of B-type reactions. Depending on the mechanism of occurrence, it can be divided into type 1-4 hypersensitivity reactions, which appear in various ways depending on the drug, and the same drug can cause hypersensitivity reactions of different mechanisms depending on the patient. For example, cephalosporin, which is a widely used antibiotic, can cause type 1 hypersensitivity reaction such as hives or anaphylaxis depending on the patient, and type 4 hypersensitivity reaction, exanthematous rash. Hypersensitivity reactions caused by quinolone antibiotics, which are increasingly used recently, are also frequently seen, and type 1 hypersensitivity reactions such as urticaria and severe type 4 hypersensitivity reactions such as DRESS syndrome, SJS, and TEN have also been reported, requiring attention.

In the present specification, "immediate drug hypersensitivity reaction" generally occurs within 1 hour after taking the drug, and symptoms such as urticaria, angioedema, and anaphylaxis usually appear. Immediate reactions are mainly caused by IgE antibody-mediated immune responses or drugs directly stimulating mast cells, and reactions such as urticaria, angioedema, rhinitis, airway constriction, itching, vomiting, diarrhea, and anaphylaxis appear alone or in combination. .

In the present specification, "delayed drug hypersensitivity reaction" occurs several days to several weeks after drug administration, and is a mechanism by activation of the complement system, formation of antigen-antibody complexes, and activation of immune cells, and is characterized by vasculitis, hematocrit abnormality, rash (fixed drug eruption, Exfoliative dermatitis, acute generalized exanthematous pustulosis (AGEP), DRESS syndrome, Stevens-Johnson syndrome (SJS), toxic epidermal necrolysis (TEN), arthralgia, proteinuria, lymphadenopathy, etc. are observed. It occurs between 1 and 8 weeks after administration, but may appear immediately after administration in patients who have previously had mild symptoms due to sensitization.

In the present specification, the drug hypersensitivity reaction may be one or more selected from the group consisting of anaphylaxis, angioedema, urticaria, macular rash, DRESS syndrome, Stevens-Johnson syndrome, toxic epidermal necrosis, and fixed drug eruption, but is not limited thereto. .

In the present specification, the drug may be at least one selected from the group consisting of NSAIDs (including aspirin), acetaminophen, cephalosporins, antibiotics, radiocontrast agents, chemotherapeutic agents, H2 blockers, allopurinol, and anticonvulsants, but are limited thereto It is not.

In the present specification, the NSAIDS is acemethacin, acetylsalicylic acid, bupexamax, diclofenac, diclofenac-sodium, diflunisal, dipyrone (metamisole), metamizole-sodium, ethenzamide, etofenamate, flufenamic acid , flurbiprofen, ibuprofen, indomethacin, isoxicam, kebuzone, ketoprofen, ketorolac, lonazolac, lornoxicam, meclofenamic acid, mefenamic acid, mofebutazone, nabumetone, Naproxen, (+)-ibuprofen, (-)-ibuprofen, (+)-naproxen, niflumic acid, oxaprozin, oxyfenbutazone, phenylbutazone, piroxicam, propipenazone, salicylamide, sulindac , tenoxicam, thiaprofenic acid, SC560, may be one or more selected from the group consisting of sulfasalazine and tolmetin, but is not limited thereto.

In the present specification, the radiographic contrast agents include Iohexo, Iopamidol, Ioversol, Iodixanol, Iopromide, Ioxaglate, and It may be one or more selected from the group consisting of diatrizoate, but is not limited thereto.

In the present specification, the antibiotic may be a β-lactam antibiotic (penicillin or cephalosporin), but is not limited thereto.

In the present specification, the drug hypersensitivity reaction may have occurred in Koreans, but is not limited thereto.

As used herein, the term “diagnosis” refers to determining a subject's susceptibility to a specific disease or disorder, determining whether a subject currently has a specific disease or disorder, or suffering from a specific disease or disorder It is a concept that includes both determining the prognosis of an object, or therametrics (eg, monitoring the condition of an object to provide information about treatment efficacy). In the present specification, the diagnosis may preferably mean early diagnosis or onset prediction, but is not limited thereto.

In this specification, the diagnosis means diagnosis of drug hypersensitivity reaction. The drug hypersensitivity reaction in the present invention may preferably be a hereditary drug hypersensitivity reaction caused by a genetic factor, and the symptoms may be intensified due to the addition of environmental influences to the genetic factor. .

In this specification, a 'primer' is a short single-stranded oligonucleotide that serves as a starting point for DNA synthesis. A primer specifically binds to a polynucleotide, which is a template, in an appropriate buffer and temperature conditions, and DNA polymerase adds a nucleoside triphosphate having a base complementary to the template DNA to the primer and connects the DNA. is synthesized Primers generally consist of 15 to 30 nucleotide sequences, and the melting temperature (Tm) of binding to the template strand varies depending on the nucleotide composition and length. The sequence of the primer does not have to have a sequence completely complementary to a part of the base sequence of the template, and it is sufficient to have sufficient complementarity within a range capable of hybridizing with the template and performing the specific function of the primer. Therefore, in the present invention, a primer pair, which is a detection agent according to the present invention, can be easily designed by referring to the nucleotide sequence of the cDNA or genomic DNA of the SNP site. Primers for detecting SNPs do not have to have sequences that are perfectly complementary to each gene sequence, as long as they have a length and complementarity suitable for the purpose of measuring the amount of mRNA by amplifying a specific section of mRNA or cDNA through DNA synthesis. Suffice. The primers for the amplification reaction are composed of a set (pair) that complementarily binds to the template (or sense) and the opposite side (antisense) of both ends of a specific section of the mRNA to be amplified.

In the present specification, 'probe' refers to RNA or DNA with a length of several to several hundred base pairs that can specifically bind to mRNA, cDNA (complementary DNA), DNA, etc. of a specific gene. It refers to a fragment of a polynucleotide, and is labeled so that the presence or absence of target mRNA or cDNA to be bound and the expression level can be confirmed. Probe selection and hybridization conditions can be appropriately selected according to techniques known in the art. The probe may be used in a diagnostic method for detecting an allele (or allele). The diagnostic methods include detection methods based on hybridization of nucleic acids, such as Southern blotting, etc., and may be provided in a form pre-bound to a substrate of a DNA chip in a method using a DNA chip. In the present invention, the primer or probe may be chemically synthesized using a phosphoramidite solid support synthesis method or other well-known methods.

In the present specification, the primer or probe may be chemically synthesized using a phosphoramidite solid support synthesis method or other well-known methods. In addition, primers or probes may be modified in various ways according to methods known in the art to the extent that hybridization with a target polynucleotide to be detected is not hindered. Examples of such modifications are methylation, capping, substitution of one or more natural nucleotides with homologs, and modifications between nucleotides, such as uncharged linkages such as methyl phosphonates, phosphotriesters, phosphoroamidates, carbamates, etc. ) or charged linkages (eg, phosphorothioate, phosphorodithioate, etc.), and binding of fluorescent or enzymatic labeling materials.

In addition, the present invention provides a kit for diagnosing drug hypersensitivity including the composition.

In the present specification, the kit may be an RT-PCR kit or a microarray chip kit, but is not limited thereto.

The kit can predict or early diagnose the onset of drug hypersensitivity by confirming SNP markers, which are diagnostic markers of drug hypersensitivity, through amplification, or by checking the expression level of SNP markers and mRNA expression levels.

The RT-PCR kit may include each primer pair capable of amplifying the nucleic acid containing the SNP site, and other than a test tube or other appropriate container, reaction buffer, deoxynucleotides (dNTPs), Taq-polymerization enzymes such as enzymes and reverse transcriptase, DNase, RNAse inhibitors, DEPC-water, sterile water, and the like. In addition, a primer pair specific to a gene used as a quantitative control may be included.

The microarray chip kit may include a microarray having a substrate on which a nucleic acid including the SNP site is immobilized. The microarray may be composed of a conventional microarray except for including the polynucleotide, primer or probe of the present invention. Hybridization of nucleic acids on microarrays and detection of hybridization results are well known in the art. The detection is, for example, by labeling a nucleic acid sample with a fluorescent material, for example, a label material capable of generating a detectable signal including materials such as Cy3 and Cy5, followed by hybridization on a microarray and the labeling material. A hybridization result can be detected by detecting a signal arising from

In the present invention, the term "individual" or "subject" means a subject for prediction or diagnosis of onset.

In the present invention, "sample" can be used without limitation as long as it is collected from a subject to be predicted or diagnosed, and for example, cells or tissues obtained by biopsy, blood, whole blood, serum, plasma, saliva, cerebrospinal fluid, various secretions, It can be urine, feces, etc. Preferably, it may be selected from the group consisting of blood, plasma, serum, saliva, nasal fluid, sputum, ascites, vaginal secretion, and urine, and may preferably be blood, plasma, or serum. The sample may be pretreated prior to use for detection or diagnosis. For example, it may include homogenization, filtration, distillation, extraction, concentration, inactivation of interfering components, addition of reagents, and the like.

또한, 본 발명은 피검체로부터 수득한 시료 중의 SNP 부위의 염기서열에 dbSNP database rs35372932, rs79736818, rs3129871, rs9557291, rs11724428, rs11123552, rs62124613, rs77389536, rs206887084, rs9557291, rs372025, rs8058114, rs143395859, rs57833801, rs139830888, rs9384423, rs6097064, rs2175562, rs3924164, rs11206477, rs62225078, rs73202933, rs72622187, rs139141104, rs9994092, rs1762852884, rs2335545, rs9688895, rs144130219, rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, 및 rs28717999으로 이루어진 Provided is a method for providing information necessary for diagnosing a drug hypersensitivity reaction comprising determining that a subject has a drug hypersensitivity reaction when one or more single nucleotide polymorphisms selected from the group exist.

또한, 본 발명은 피검체로부터 수득한 시료 중의 SNP 부위의 염기서열에 dbSNP database rs35372932, rs79736818, rs3129871, rs9557291, rs11724428, rs11123552, rs62124613, rs77389536, rs206887084, rs9557291, rs372025, rs8058114, rs143395859, rs57833801, rs139830888, rs9384423, rs6097064, rs2175562, rs3924164, rs11206477, rs62225078, rs73202933, rs72622187, rs139141104, rs9994092, rs1762852884, rs2335545, rs9688895, rs144130219, rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, 및 rs28717999으로 이루어진 Provided is a method for providing information necessary for predicting the onset of a drug hypersensitivity reaction, comprising determining that the subject has a high risk of drug hypersensitivity reaction when one or more single nucleotide polymorphisms selected from the group exist.

In addition, the present invention dbSNP database rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, and rs2985 selected from a single group consisting of rs1276175, rs144157017, rs59594955, and rs2987 Provided is a method for providing information necessary for diagnosis of a delayed drug hypersensitivity reaction, comprising determining that the subject has a delayed drug hypersensitivity reaction when the polymorphism exists.

In the present specification, the method includes sequencing, exome sequencing, microarray hybridization, allele specific PCR, and dynamic allele-specific hybridization. hybridization), PCR extension analysis, and one or more methods selected from the group consisting of Taqman technique, but is not limited thereto.

Since the present invention can apply various transformations and have various embodiments, specific embodiments will be illustrated in the drawings and described in detail in the detailed description. However, this is not intended to limit the present invention to specific embodiments, and should be understood to include all conversions, equivalents, or substitutes included in the spirit and scope of the present invention. In describing the present invention, if it is determined that a detailed description of related known technologies may obscure the gist of the present invention, the detailed description will be omitted.

Hereinafter, a preferred embodiment is presented to aid understanding of the present invention. However, the following examples are provided to more easily understand the present invention, and the content of the present invention is not limited by the following examples.

[Example]

Example 1. Experimental method

Target

From 2012 to 2017, 378 patients with DHR (Drug Hypersensitivity Reactions) who visited Asan Medical Center were recruited. This study was conducted in accordance with the Declaration of Helsinki and the protocol was approved by the Institutional Review Board of Asan Medical Center (2011-0939). Written informed consent was obtained from all participants for enrollment and blood sample collection. Blood samples were collected from each subject and genotyped using K-CHIP. K-CHIP was provided through the K-CHIP consortium (http://kogo.or.kr) and was designed by the Center for Genome Science at the Korea Institute of Health. Control data (N = 3,780) used the data of the domestic Severance Hospital Health Examination Center, and genotype was analyzed using K-CHIP (see Fig. 1). SNP genotypes were analyzed using a K-medoid clustering-based calling method to minimize batch effects. The EIGENSTRAT approach was applied to the pooled data and principal component (PC) scores were calculated between control and patient groups (to match subjects with similar genetic information). A total of 10 controls per patient were matched with similar scores. The PC score was used to identify subjects with no genetic background. A multidimensional scaling plot is shown in FIG. 2 .

Replication analysis

Genotyping was repeated for 185 additional DHR patients recruited from Asan Medical Center between 2012 and 2015. These patients were genotyped using an Axiom Asian chip (Affymetrix). The control group was enrolled at the Severance Hospital Health Examination Center (N=2,312) and genotype was analyzed using K-CHIP. All data were pooled and object imputation was performed (see Figure 1).

data analysis group

DHR patients were divided into two groups: immediate response and delayed response. For comparative analysis, total DHR was compared with the control group, each DHR group was compared with the control group, and finally, the immediate and delayed response groups were compared.

Quality Management

The SNP and the quality of the subjects, that is, the quality was evaluated. All steps of quality control were performed using PLINK and ONETOOL. SNPs with a P-value of less than 1 × 10 ^-5 for the Hardy-Weinberg equilibrium (HWE) test, a minor allele frequency (MAF) of less than 0.05, or a genotyping rate of less than 0.95 were excluded. In addition, individuals with a missing rate greater than 0.05 or IBS (Identity-By-State) greater than 0.9 along with other individuals were excluded. This filtering procedure was applied to each case and control separately, and cases and controls were combined. In addition, SNPs present in both groups were considered, and the same quality control procedures were applied to the pooled data. In addition, Fisher's exact test was used to test the equivalence of average missing rates between each case and control group, and significant SNPs at the 1 × 10 ^-5 significance level were removed from the GWAS. A summary of this process is shown in Figure 1A.

Statistical analysis using genotypic imputation and inferred genotypes

GWAS with discovery data were performed using generalized logistic regression (GLR). Age and gender were included as covariates. In addition, a logistic regression (LR) model was used to predict a binary response (control versus DHR). Both GLR and LR were applied using R statistical software (R Project for Statistical Computing, Vienna, Austria; www.r-project.org). Genome-wide significant SNPs were selected at the 5 × 10 ^-8 significance level. For fine mapping, SNPs located within ± 5,000 base pairs of significant SNPs in the whole genome were inferred. Inference was performed using the Sanger inference service (https://imputation.sanger.ac.uk). Haplotype Reference Consortium release v 1.1 was used and Asians of European ancestry (CHB/JPT)16 consisting of 64,940 haplotypes were considered as the main reference panel. Pre-phasing and inference were performed using SHAPEIT17 and PBWT18, and inference accuracy was evaluated using the INFO metric.

Example 1. Confirmation of subject characteristics

The demographic characteristics of study participants are summarized in Table 1.

The mean age of DHR patients (N=378) enrolled in the study was 42.03 ± 14.82 years, and 247 patients (65.34%) were female. Of these, 295 cases (78.04%) showed immediate hypersensitivity reactions and 83 cases (21.96%) showed delayed/non-immediate reactions. The most common drugs causing immediate hypersensitivity reactions were NSAIDs, followed by antibiotics. A detailed list of causative drugs is shown in Table 2 below.

In the delayed response group, antibiotics were the most common causative drug, followed by anticonvulsants. The mean age of the control group (N=3,779) was 43.07 ± 9.85 years, and a total of 1236 patients (32.7%) were female. In the replication analysis, the mean age of the 185 DHR patients enrolled was 50.38 ± 16.58 years, and 105 women (56.76%) were included. Of the 185 patients, 80 patients showed immediate-type hypersensitivity reactions, whereas 105 patients showed delayed-type hypersensitivity reactions. The control group consisted of 2,312 subjects, including a total of 921 women (39.84%), and the average age of the subjects was 46.96±10.72 years. In the replication group, NSAIDs were the most common causative drug, followed by antibiotics (Table 2).

characteristic drug hypersensitivity Control discovery data N=378 N=3,780 age (years) 42.03 ± 14.82 43.07 ± 9.85 female, N (%) 247 (65.34%) 1236 (32.70%) drug hypersensitivity Immediate, N (%) 295 (78.04%) Anaphylaxis 147 (49.83%) angioedema 54 (18.30%) Urticaria ± Angioedema 94 (31.86%) Delayed form, N (%) 83 (21.96%) maculopapular rash 36 (43.37%) DRESS 20 (24.10%) Stevens-Johnson Syndrome 13 (15.66%) toxic epidermal necrosis 10 (12.05%) fixed advance 4 (4.82%) duplicate data N=185 N=2,312 age (years) 50.38 ± 16.58 46.96 ± 10.72 female, N (%) 102 (55.14%) 921 (39.84%) drug hypersensitivity Immediate, N (%) 105 (56.76%) Anaphylaxis 55 (52.38%) angioedema 4 (3.81%) Urticaria ± Angioedema 46 (43.81%) Delayed form, N (%) 80 (43.24%) maculopapular rash 41 (51.25%) DRESS 24 (30.00%) Stevens-Johnson Syndrome 8 (10.00%) toxic epidermal necrosis 5 (6.25%) fixed advance 1 (1.25%) Acute generalized exanthematous pustulosis 1 (1.25%)

discovery immediate response (N=295) Delayed response (N=83) NSAIDs (including aspirin) 125 7 acetaminophen 12 3 cephalosporin 75 8 other antibiotics 19 27 radiocontrast agent 11 5 chemotherapeutic agent 6 2 H2 blockers 32 One allopurinol One 6 anticonvulsants 0 13 etc 14 11 a copy immediate response (N=105) Delayed response (N=80) NSAIDs (including aspirin) 48 3 acetaminophen 2 One cephalosporin 17 21 other antibiotics 18 23 radiocontrast agent 12 One chemotherapeutic agent One One H2 blockers 5 One allopurinol 0 10 anticonvulsants 0 10 etc 2 9

Example 2. Confirmation of GWAS analysis results

A total of 265,878 SNPs passed quality control (call rate > 98%, MAF > 0.05). First, the association between the two groups of DHR (immediate and delayed response) was analyzed. The Q-Q plot for LR is shown in Figure 3, and the analysis results were statistically significant.

On the other hand, as shown in FIG. 4, only one SNP appeared close to the genome-wide significance level (rs35372932, p = 7.99E-08). The genome-wide SNP is rs35372932, which was confirmed to be present in the upstream region of the HLA-DRB1 gene on chromosome 6 (see Table 3).

In addition, as shown in Figure 5a, a region plot using genotypes and alternative SNPs, purple filled dots represent rs35372932, and other dots' colors represent correlations with rs35372932. That is, the most significant SNP was rs35372932.

Next, the analysis was repeated by dividing the immediate and delayed responses independently. Q-Q plots for GLR are shown in Figures 3b and 3c.

Meanwhile, 5 significant SNPs (rs11206477, rs62225078, rs73202933, rs72622187 and rs139141104) on the whole genome were compared between the delayed hypersensitivity reaction and the control group (see Tables 3 and 4).

As shown in Table 3 above, the most significant SNP rs139141104 is present in the intron region of the MUC22 gene of chromosome 6.

As shown in FIG. 6, it was confirmed that a total of 5 SNPs (rs11206477, rs62225078, rs73202933, rs72622187 and rs139141104) were significant in the full-length gene. This is the same result as shown in Table 4.

On the other hand, in the area plot of FIG. 5B prepared using genotypes and alternative SNPs, the purple filled dots represent rs139141104, and the colors of the other dots represent their correlations.

Finally, we compared the phenotypes of drug hypersensitivity reactions.

As shown in Table 4, when comparing immediate and delayed hypersensitivity reactions, no significant SNPs were identified.

rsID BP Alt/Ref MAF _ALL MAF_
_con MAF_
_immediate MAF_
_delayed MAF
_{_CODA} MAF
_{_gnomAD-east asian} HWE Gene P-value Model 1 (control vs DHR) rs35372932 32597208 T/C 0.287 0.278 0.37 0.398 0.2692 0.285 0.91 HLA-DRB1 7.99E-08 rs79736818 148357581 G/A 0.05521 0.05184 0.08644 0.09756 - 0.04058 0.3272 LOC102546294 6.44E-06 rs3129871 32438565 A/C 0.3111 0.318 0.2356 0.2683 - 0.7017 0.1548 HLA-DRA 6.67E-06 rs9557291 99761210 A/G 0.05738 0.05383 0.1017 0.06098 - 0.06919 0.6459 CLYBL 1.03E-05 rs11724428 112598451 A/G 0.384 0.3767 0.461 0.4451 - 0.4506 0.3426 ZGRF1 1.13E-05 - 141452237 C/T 0.1547 0.16 0.09661 0.1159 - - 0.6544 - 1.56E-05 rs11123552 120454245 G/C 0.2158 0.2103 0.2712 0.2683 - 0.2394 0.3846 - 1.75E-05 rs62124613 20386466 T/C 0.06798 0.06417 0.1068 0.1037 - 0.08956 0.1686 - 1.78E-05 rs77389536 102408368 C/T 0.2584 0.2523 0.3102 0.3537 - 0.2977 0.6965 PSD 1.80E-05 rs206887084 94620980 A/G 0.08703 0.08313 0.122 0.1402 - - 0.387 - 1.92E-05 Model2 (control vs immediate) rs9557291 99761210 A/G 0.05738 0.05383 0.1017 0.06098 - 0.07226 0.5522 CLYBL 1.05E-06 rs372025 107077214 C/T 0.185 0.1806 0.2559 0.1341 - 0.7642 0.5712 - 1.08E-06 rs8058114 12603772 A/G 0.07196 0.06921 0.1186 0.03049 - 0.03535 0.09014 - 2.65E-06 rs143395859 27293404 A/G 0.111 0.1069 0.1627 0.1159 - 0.1247 0.2229 - 8.22E-06 rs57833801 121777405 A/G 0.4562 0.4487 0.4525 0.4756 - 0.4239 0.02729 LOC105373590 1.03E-05 rs139830888 147872742 A/G 0.06931 0.06603 0.1169 0.04878 0.07106 0.05962 0.9046 CNTNAP2 1.05E-05 rs9384423 156089071 G/A 0.1322 0.1277 0.1915 0.128 - 0.1615 0.387 LOC101928923 1.18E-05 rs6097064 37952348 C/A 0.3862 0.3803 0.4678 0.3659 - - 0.4857 - 1.48E-05 rs2175562 140398639 G/A 0.4488 0.4427 0.4746 0.4512 - 0.5335 0.2026 CLGN 1.53E-05 rs3924164 76737927 T/C 0.4859 0.4798 0.4322 0.4695 - 0.4008 31010 - 1.74E-05 Model2 (control vs delayed) rs11206477 54941621 A/C 0.05123 0.05251 0.04915 0 - 0.03077 0.03531 - <1.00E-10 rs62225078 47740975 A/T 0.06027 0.06059 0.07288 0 - 0.1123 0.5396 LOC284930 <1.00E-10 rs73202933 89819306 C/T 0.05304 0.05357 0.06102 0 - 0.04231 0.8757 - <1.00E-10 rs72622187 6078051 A/G 0.06811 0.06828 0.08475 0 - 0.1662 0.2273 LOC105376942 <1.00E-10 rs139141104 31021244 G/A 0.08715 0.08831 0.08305 0.04878 - 0.008366 0.3287 MUC22 2.90E-08 rs9994092 65570396 A/C 0.05268 0.05145 0.04237 0.1463 - 0.04557 0.7668 EPHA5 2.65E-07 rs1762852884 30994936 C/A 0.06931 0.06988 0.04915 0.1159 - - 0.8768 MUC22 9.31E-07 rs2335545 2916111 T/C 0.2183 0.2153 0.2136 0.372 - 0.2365 0.2096 FLYWCH1 1.51E-06 rs9688895 38465523 A/C 0.08595 0.08446 0.07797 0.1829 - 0.0906 0.2557 BTBD9 1.54E-06 rs144130219 98725782 C/T 0.05219 0.05078 0.04746 0.1341 - 0.1004 0.002304 SLC15A1 1.74E-06 Model 3 (Immediate vs Delayed) rs78831487 6537140 C/T 0.1084 0.08136 0.2073 - 0.1223 One - 2.08E-06 rs456449 38851003 A/G 0.05304 0.0339 0.128 - 0.0739 0.7297 - 6.66E-06 rs13250548 35650882 G/A 0.2435 0.2102 0.3963 - 0.2442 0.5583 - 7.00E-06 rs77068773 23204196 T/A 0.1262 0.1034 0.2073 - 0.1538 0.07611 - 1.78E-05 rs3025035 43783622 T/C 0.2108 0.2017 0.3476 0.2032 0.1532 0.05472 VEGFA 2.44E-05 rs2088142 33691285 A/G 0.429 0.3881 0.4268 - 0.4238 0.9328 RYR3 2.46E-05 rs1276175 30945691 A/G 0.4461 0.4254 0.4329 - 0.42377 0.3581 2.52E-05 rs144157017 50740465 G/A 0.05617 0.03051 0.1037 - 0.05591 0.2289 TFAP2D 2.59E-05 rs59594955 38443346 C/A 0.08691 0.07797 0.1829 - 0.08805 0.231 BTBD9 2.82E-05 rs28717999 65571223 G/A 0.05243 0.04068 0.1402 - 0.0482 0.4389 EPHA5 3.02E-05

Example 3. Replication analysis

Replication analysis was performed using replacement. A total of 75,077 SNPs passed quality control (genotyping rate > 98%, MAF > 0.03) (FIG. 1).

As shown in Table 5, among the SNPs identified as significant in the discovery data analysis, only rs35372932 (p=0.008), an HLA-DRB1 upstream region of chromosome 6, was statistically significant. This is a result supporting that rs35372932 can be used as a universal marker that is not limited to a specific drug hypersensitivity phenotype or causative drug.

Example 4. GCTA (Genome-wide Complex Trait Analysis) analysis

GCTA is one of the methods for estimating heritability, and one of the methods for estimating the heritability of SNPs.

The present inventors calculated the ratio of phenotypic variation using hSNP ² (additive SNP heritability) of genotypic SNPs. Results presented estimates of hSNP ² according to prevalence.

As shown in FIG. 7 , as a result of setting the prevalence of drug hypersensitivity reaction to 2 × 10 ^-2 , hSNP ² was found to be 39.12% in the responsibility scale. This is a result that supports the fact that drug hypersensitivity has a relatively high heritability, in light of previous studies.

According to the above examples, SNPs judged to have clinical significance in drug hypersensitivity and its detailed types, immediate drug hypersensitivity and delayed drug hypersensitivity, were selected. The SNPs of the present invention are used for early diagnosis and It is expected that it will be useful for disease prediction.

The above description of the present invention is for illustrative purposes, and those skilled in the art can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, the embodiments described above should be understood as illustrative in all respects and not limiting.

<110> THE ASAN FOUNDATION University of Ulsan Foundation For Industry Cooperation <120> SNP for drug hypersensitivity and diagnosis method using the same <130> MP21-119 <160> 38 <170> KoPatentIn 3.0 <210> 1 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs35372932 <220> <221> variation <222> (11) <223> r = C or T <400> 1 cagtatcata cagcagtttt g 21 <210> 2 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs79736818 <220> <221> variation <222> (11) <223> r = A or G <400> 2 aagccttctc ataggtccgc a 21 <210> 3 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs3129871 <220> <221> variation <222> (11) <223> r = A or G <400> 3 gtgggaaagaa attaacattc t 21 <210> 4 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs9557291 <220> <221> variation <222> (11) <223> r = G or A <400> 4 aaaattagcc gggcgtggtg g 21 <210> 5 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs11724428 <220> <221> variation <222> (11) <223> r = G or A <400> 5 ttataataac gaaaacattg a 21 <210> 6 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs11123552 <220> <221> variation <222> (11) <223> r = C or G <400> 6 ccactgcact ccagcctggg t 21 <210> 7 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs62124613 <220> <221> variation <222> (11) <223> r = C or G <400> 7 gctgaggctg cgaagggaga t 21 <210> 8 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs77389536 <220> <221> variation <222> (11) <223> r = T or C <400> 8 gggcacacac tgttcacacc c 21 <210> 9 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs9557291 <220> <221> variation <222> (11) <223> r = G or A <400> 9 aaaattagcc gggcgtggtg g 21 <210> 10 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs372025 <220> <221> variation <222> (11) <223> r = T or C <400> 10 ttcattcttt ttaagtcgaa g 21 <210> 11 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs8058114 <220> <221> variation <222> (11) <223> r = G or A <400> 11 ttgcagtgac ggcatcaggt g 21 <210> 12 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs143395859 <220> <221> variation <222> (11) <223> r = G or A <400> 12 attgatttgc gtatgttgaa c 21 <210> 13 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs57833801 <220> <221> variation <222> (11) <223> r = G or A <400> 13 attacatggg gccaagggag t 21 <210> 14 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs139830888 <220> <221> variation <222> (11) <223> r = G or A <400> 14 ttttgggggc gtagattact t 21 <210> 15 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs9384423 <400> 15 aagcacccag aactagatgg a 21 <210> 16 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs6097064 <220> <221> variation <222> (11) <223> r = A or C <400> 16 gtaattccag aactttggga g 21 <210> 17 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs2175562 <220> <221> variation <222> (11) <223> r = A or G <400> 17 aatatatatt aaaaaatgca a 21 <210> 18 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs3924164 <220> <221> variation <222> (11) <223> r = C or T <400> 18 gggcatatct cgagcaagga a 21 <210> 19 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs11206477 <220> <221> variation <222> (11) <223> r = C or A <400> 19 ccacaaattt ctcttccttt c 21 <210> 20 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs62225078 <220> <221> variation <222> (11) <223> r = T or A <400> 20 agcaatgcag ttttcccttc c 21 <210> 21 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs73202933 <220> <221> variation <222> (11) <223> r = T or C <400> 21 gtatctatta tattaatcca c 21 <210> 22 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs72622187 <220> <221> variation <222> (11) <223> r = G or A <400> 22 aagtaaaaga ggggggcaaa a 21 <210> 23 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs139141104 <220> <221> variation <222> (11) <223> r = A or G <400> 23 aaatacacca atcagcaccc t 21 <210> 24 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs9994092 <220> <221> variation <222> (11) <223> r = C or A <400> 24 ttagtttttg ctatggttgg t 21 <210> 25 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs1762852884 <220> <221> variation <222> (11) <223> r = A or C <400> 25 ctgaggtgac cacagtcttt g 21 <210> 26 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs2335545 <220> <221> variation <222> (11) <223> r = C or T <400> 26 ttttgggagg caaagacggg c 21 <210> 27 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs9688895 <220> <221> variation <222> (11) <223> r = C or A <400> 27 cccagctact cgggaggctg a 21 <210> 28 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs144130219 <220> <221> variation <222> (11) <223> r = T or C <400> 28 tggggtgcag tgatgtgatc a 21 <210> 29 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs78831487 <220> <221> variation <222> (11) <223> r = T or C <400> 29 aggctggtct tgaactccga g 21 <210> 30 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs456449 <220> <221> variation <222> (11) <223> r = G or A <400> 30 ttggttcttc ggccttttgc g 21 <210> 31 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs13250548 <220> <221> variation <222> (11) <223> r = A or G <400> 31 atggttttca atgagaatga g 21 <210> 32 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs77068773 <220> <221> variation <222> (11) <223> r = A or T <400> 32 tgtgaccttc aattaattat a 21 <210> 33 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs3025035 <220> <221> variation <222> (11) <223> r = C or T <400> 33 ctctaacctc ccaatacctt t 21 <210> 34 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs2088142 <220> <221> variation <222> (11) <223> r = G or A <400> 34 ttattggttc gttgagttaa t 21 <210> 35 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs1276175 <220> <221> variation <222> (11) <223> r = G or A <400> 35 gtctcactcc atcacccagg t 21 <210> 36 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs144157017 <220> <221> variation <222> (11) <223> r = A or G <400> 36 atgtagtctc actttgtcgc c 21 <210> 37 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs59594955 <220> <221> variation <222> (11) <223> r = A or C <400> 37 ggctcaactt accacacctc a 21 <210> 38 <211> 21 <212> DNA <213> artificial sequence <220> <223> rs28717999 <220> <221> variation <222> (11) <223> r = A or G <400> 38 agtagggggg actatatctt a 21

Claims (13)

dbSNP 데이터베이스 rs35372932, rs79736818, rs3129871, rs9557291, rs11724428, rs11123552, rs62124613, rs77389536, rs206887084, rs9557291, rs372025, rs8058114, rs143395859, rs57833801, rs139830888, rs9384423, rs6097064, rs2175562, rs3924164, rs11206477, rs62225078, rs73202933, rs72622187, rs139141104, rs9994092, rs1762852884, rs2335545, rs9688895, rs144130219, rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, 및 rs28717999로 이루어진 군으로부터 선택된 하나 이상의 SNP의 검출 제제를 포함하는 약물 과민반응의 진단 또는 A composition for predicting onset.
According to claim 1,
The drug hypersensitivity reaction is an immediate drug hypersensitivity reaction or a delayed drug hypersensitivity reaction, characterized in that, a composition for diagnosing or predicting the onset of drug hypersensitivity reaction.
According to claim 1,
If the SNP is one or more selected from the group consisting of i) rs35372932, rs79736818, rs3129871, rs9557291, rs11724428, rs11123552, rs62124613, rs77389536, and rs206887084, the composition is for diagnosing or predicting the onset of drug hypersensitivity reactions from a normal person,
If the SNP is one or more selected from the group consisting of ii) rs9557291, rs372025, rs8058114, rs143395859, rs57833801, rs139830888, rs9384423, rs6097064, rs2175562, and rs3924164 for diagnosis or predicting normal onset drug, the composition is is,
If the SNP is one selected from the group consisting of: iii) rs11206477, rs62225078, rs73202933, rs72622187, rs139141104, rs9994092, rs1762852884, rs2335545, rs9688895, and rs144130219 for predicting a hypersensitivity-onset reaction or abnormal drug, the composition for normal diagnosis or abnormality is,
If the SNP is at least one selected from the group consisting of iv) rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, and rs28717999, the composition for distinguishing between immediate drug hypersensitivity reaction and delayed drug hypersensitivity reaction is at least one composition, Characterized in that, a composition for diagnosing or predicting the onset of drug hypersensitivity reactions.
According to claim 1,
The SNP is rs35372932 or rs139141104, characterized in that, a composition for diagnosing or predicting the onset of drug hypersensitivity reactions.
According to claim 1,
The drug hypersensitivity reaction is a composition for diagnosing or predicting the onset of drug hypersensitivity reaction, characterized in that it has occurred in Koreans.
According to claim 1,
The detection agent is a composition for diagnosing or predicting the onset of drug hypersensitivity, characterized in that the probe capable of detecting the SNP of claim 1.
According to claim 1,
The detection agent is a composition for diagnosing or predicting the onset of drug hypersensitivity, characterized in that the primer capable of detecting the SNP of claim 1.
A kit for diagnosing a drug hypersensitivity reaction comprising the composition of claim 1.
According to claim 8,
Characterized in that the kit is an RT-PCR kit or a microarray chip kit.
피검체로부터 수득한 시료 중의 SNP 부위의 염기서열에 dbSNP database rs35372932, rs79736818, rs3129871, rs9557291, rs11724428, rs11123552, rs62124613, rs77389536, rs206887084, rs9557291, rs372025, rs8058114, rs143395859, rs57833801, rs139830888, rs9384423, rs6097064, rs2175562 , rs3924164, rs11206477, rs62225078, rs73202933, rs72622187, rs139141104, rs9994092, rs1762852884, rs2335545, rs9688895, rs144130219, rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, 및 rs28717999으로 이루어진 군으로부터 선택된 하나 이상의 A method for providing information necessary for diagnosing a drug hypersensitivity reaction comprising determining that a subject suffers from a drug hypersensitivity reaction when a single nucleotide polymorphism exists.
피검체로부터 수득한 시료 중의 SNP 부위의 염기서열에 dbSNP database rs35372932, rs79736818, rs3129871, rs9557291, rs11724428, rs11123552, rs62124613, rs77389536, rs206887084, rs9557291, rs372025, rs8058114, rs143395859, rs57833801, rs139830888, rs9384423, rs6097064, rs2175562 , rs3924164, rs11206477, rs62225078, rs73202933, rs72622187, rs139141104, rs9994092, rs1762852884, rs2335545, rs9688895, rs144130219, rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, 및 rs28717999으로 이루어진 군으로부터 선택된 하나 이상의 A method for providing information necessary for predicting the onset of a drug hypersensitivity reaction comprising determining that a subject has a high risk of developing a drug hypersensitivity reaction when a single nucleotide polymorphism exists.
A dbSNP database rs78831487, rs456449, rs13250548, rs77068773, rs3025035, rs2088142, rs1276175, rs144157017, rs59594955, and rs2871799 consisting of one or more single salts present in the base sequence of the SNP site in the sample obtained from the patient with drug hypersensitivity reaction In this case, a method for providing information necessary for diagnosis of a delayed drug hypersensitivity reaction comprising determining that the subject has a delayed drug hypersensitivity reaction.
According to any one of claims 10 to 13,
The method includes sequencing, exome sequencing, microarray hybridization, allele specific PCR, dynamic allele-specific hybridization, PCR Characterized in that it is performed by at least one method selected from the group consisting of extension analysis and Taqman technique.

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