KR20210020253A - Multiplex drug gene diagnosis kit for predicting drug side effects on chronic diseases and cancer related multi-prescription drugs and personalized drug treatment - Google Patents

Abstract

The present invention relates to a kit for multiple high-speed drug gene analysis for predicting side effects of a drug and personalized drug treatment for multi-prescription drugs related to cancer and chronic diseases. According to the present invention, the kit for multiple high-speed drug gene analysis comprises primers of SEQ ID NOS: 1 to 124, and comprises shrimp alkaline phosphatase (SAP) and/or exonuclease 1. According to the present invention, side effects of a drug can be rapidly and conveniently diagnosed.

Description

Multiple drug gene diagnosis kit for predicting drug side effects on chronic diseases and cancer related multi-prescription drugs and personalized drug treatment}

The present invention relates to a kit for diagnosing multiple high-speed drug genes for predicting drug side effects and personalized drug treatment for cancer and chronic disease-related multi-prescription drugs. More specifically, ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1,*2,*3,*17, CYP2C9 *1,*2,*3,*13,*14 , CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN (duplicate), CYP4F2 *1 ,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, KCNJ11 rs5219, MMP3 rs3041425058, NEDT rs15 rs5219, 232,DD4 PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A, *1B, *5, *15, TPMT *1, *3C, *6, UGT1A1 *1, *28, VKORC1 rs9923231 A total of 27 genes of 50 specific mutations A kit for quickly determining the presence or absence using tSNP, and a method of diagnosing multiple high-speed drug genes for personalized drug therapy and predicting drug side effects for cancer and chronic disease-related multi-prescription drugs using the same. Therefore, it relates to the use of evaluating individual differences in drug reactions and the risk of adverse reactions.

The content described below merely provides background information related to the present invention and does not constitute the prior art.

According to the British medical journal, among the deaths due to adverse drug reactions, less than 11% of the deaths caused by the doctor's prescription or dispensing process are wrong, and in most cases, unexpected drugs that occur in patients who take the drug properly according to the normal drug prescription. It is said to be due to side effects. In addition, according to a survey by the American Medical Association, more than 2 million American patients who normally take prescribed drugs experience serious adverse drug reactions each year, and more than 100,000 patients die from adverse drug reactions each year. There are also reports that the resulting economic loss is estimated to amount to $180 billion. In Korea, it has been reported that information on drug side effects reported in 2016 was 22,939, an increase of 15.6% from the previous year. These adverse drug reactions can be caused by a variety of causes, such as dietary habits, genetics, and environmental factors. Among them, genetic factors that help predict individual differences in drug reactions and risk of adverse reactions are a significant number of genetic biomarkers. It is known through research and verification.

However, since various types of gene mutations have been reported, it is important to determine the correct genotype through a minimum of single nucleotide polymorphism (SNP) in order to increase the efficiency of analysis time and cost. Recently, Roche's AmpliChip CYP450 Test, which analyzes SNPs based on CYP2D6 gene mutations mainly found in Westerners containing some of the above genes, Luminex's xTAG ^® CYP2D6 Kit v3, and Bioneer's AccuPower® Warfarin genotyping Kit products from Korea have been reported. Has become. However, since the combination of mutant genes is mainly composed of mutations found in Westerners, or only one or two drug genes can be tested, products that evaluate individual differences in drug reactions and risk of adverse reactions prescribed in Korea are very insufficient. Actually.

Accordingly, the present inventors developed a multi-fast drug gene diagnosis technology to rapidly and effectively diagnose individual differences in drug reactions and major mutations related to adverse reactions, centering on drugs that are multiprescribed for cancer and chronic diseases in Koreans, and utilize this technology. ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1,*2,*3,*17, CYP2C9 *1,*2,*3,*13,*14, CYP2D6 1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN (duplicate), CYP4F2 *1,*3 , DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs25041458, NEDD4 rs30L15rs5219, MMP3 rs30L15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*1B,*5,*15, TPMT *1,*3C,*6, UGT1A1 *1,*28, VKORC1 rs9923231 A total of 27 genes of 50 specific It was confirmed that the presence or absence of the allele can be quickly and accurately detected, and the present invention was completed.

Nelson et al., Pharmacogenetics, 6:1-42, 1996 J Natl. Cancer Inst., 83:1164-1168 Tai HL. Am J Hum Genet. 58:694-702, 1996 Ameyaw M-M. et al., Hum Mol Genet. 8:367-370, 1999 Clin Pharmacol Ther. 2018 Feb;103(2):210-216 Clin Pharmacol Ther. 2014 Oct;96(4):423-8 Clin Pharmacol Ther. 2017 Sep;102(3):397-404 Int. J. Mol Sci. 2016 Nov; 17(11):1890 Nat Genet. 2014 Sep;46(9):1017-20

An object of the present invention is to rapidly detect mutations in biomarkers related to cancer and chronic diseases, and to provide a multi-fast drug gene diagnosis kit for predicting drug side effects for cancer and chronic disease-related multi-prescription drugs and personalized drug treatment. .

Another object of the present invention is to provide a method for diagnosing multiple high-speed pharmacokinetics for predicting drug side effects and personalized drug therapy for the multi-prescription drugs using the kit.

In order to solve the above problems, the present invention, in the multi-speed drug gene diagnostic kit for predicting drug side effects and personalized drug treatment for cancer and chronic disease-related multi-prescription drugs, comprising the primers of SEQ ID NOs: 1 to 124, , ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1,*2,*3,*17, CYP2C9 *1,*2,*3,*13,*14, CYP2D6 1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN (duplicate), CYP4F2 *1,*3 , DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs25041458, NEDD4 rs30L15rs5219, MMP3 rs30L15 One selected from the group consisting of rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*1B,*5,*15, TPMT *1,*3C,*6, UGT1A1 *1,*28 and VKORC1 rs9923231 It provides a kit for diagnosing multiple high-speed pharmacological genes for predicting drug side effects and personalized drug treatment for cancer and chronic disease-related multi-prescription drugs, characterized in determining the presence or absence of the above mutations.

In the present invention, the kit consists of 2X PCR Amplification Mix, 2X Multiplex PCR Amplification Mix, amplification primer mix 1 (APM1) consisting of primers of SEQ ID NO: 1 to SEQ ID NO: 4, and primers of SEQ ID NO: 5 to SEQ ID NO: 6 Amplification primer mix 2 (APM2), amplification primer mix 3 (APM3) consisting of primers of SEQ ID NO: 7 to SEQ ID NO: 8, amplification primer mix 4 (APM4) consisting of primers of SEQ ID NO: 9 to 24, SEQ ID NO: 25 Amplification primer mix 5 (APM5) consisting of primers of SEQ ID NOs: 29-44, amplification primer mix 6 (APM6) consisting of primers of SEQ ID NOs: 29-44, amplification primer mix 7 (APM7) consisting of primers of SEQ ID NOs: 45-46 , Amplification primer mix 8 consisting of primers of SEQ ID NOs: 47 to 56 (APM8), amplification primer mix 9 (APM9) consisting of primers of SEQ ID NOs: 57 to 66, an amplification primer mix consisting of primers of SEQ ID NOs: 67 to 76 10 (APM10), snapshot multiplex reagent (SNaPshot MR), snapshot primer mix 1 consisting of primers of SEQ ID NOs: 77 to 87 (SPM1), snapshot primer mix 2 consisting of primers of SEQ ID NOs: 88 to 99 ( SPM2), the snapshot primer mix 3 (SPM3) consisting of the primers of SEQ ID NOs: 100 to 109, the snapshot primer mix 4 (SPM4) consisting of the primers of SEQ ID NOs: 110 to 119, consisting of the primers of SEQ ID NOs: 120 to 124 Snapshot Primer Mix 5 (SPM5), control DNA and nuclease-free water.

In addition, the kit may further include Shrimp Alkaline Phosphatase (SAP) and/or Exonuclease 1 (Exo).

In the present invention, the side effects may be selected from the group consisting of Stevens-Johnson syndrome, toxic epidermal necrosis dissolution, irritability syndrome, severe liver injury, and muscle disease.

In the present invention, the drug is voriconazole, piroxicam, aspirin, celecoxib, diclofenac, tramadol, ibuprofen, omeprazole, esomeprazole, lansoprazole, pantoprazole, rabeprazole, simbassatin, cerivastatin , Pravastatin, rosuvastatin, atorvastatin, clopidogrel, flecainide, propaphenone, nifedipine, captopril, quinacryl, enalapril, lisinopril, telmisartan, losartan, metoprolol, irbesartan, cande Sartan, carvedirol, hydrochlorothiazide, warfarin, glimepiride, glibenclamide, gliclazide, metformin, allopurinol, lapatinib, tamoxifen, cisplatin, mercaptopurine, pazopanib, capecitabine, fluorine It may be one or more selected from the group consisting of loracil, irinotecan, azathioprine, rasburicase, and similar drugs.

The present invention also provides a method for diagnosing multiple high-speed pharmacological genes for predicting drug side effects and personalized drug treatment for cancer and chronic disease-related multi-prescription drugs, comprising the following steps:

(a) Using the above kit, ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1, *2, *3, *17, CYP2C9 *1, *2, *3, *13,*14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN (duplicate ), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 47319T, KCNJ11 rs5219T , MMP3 rs3025058, NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*1B,*5,*15, TPMT *1,*3C,*6, UGT1A1 *1,*2 KORC Determining a SNP capable of specifically detecting at least one genotype selected from the group consisting of;

(b) ABCG2, ACE, ADRA1A, AGTR1, C11orf65, COQ2, CYP2C19, CYP2C9, CYP2D6, CYP4F2, DPYD, G6PDGNB3, HLA-B, HLA-DQA1, HLA-DRB1, Internal Control HGF, NEDD15, NUDT15, PTGS2, NUDT15 , SLC14A2, SLCO1B1, TPMT, UGT1A1, and performing a multiple polymerase chain reaction to amplify one or more genes selected from the group consisting of VKORC1; And

(c) performing a SNAPshot analysis using multiple single base primers to analyze the SNP for the product to confirm the genotype.

In the present invention, in the step (b), the multiple polymerase chain reaction may be performed using one or more primers selected from the group consisting of primers of SEQ ID NOs: 1 to 76.

Specifically, CYP2D6 was specifically amplified using SEQ ID NOs: 1 to 8, CYP2C19 was specifically amplified using SEQ ID NOs: 9 to 14, and CYP4F2 was specifically amplified using SEQ ID NOs: 15 to 16. And, using SEQ ID NOs: 17 to 22, the CYP2C9 gene is specifically amplified, and the VKORC1 gene is specifically amplified using SEQ IDs: 23 to 24, and the HLA-B gene is specific using SEQ ID NOs: 25 to 26. Amplify specifically, amplify the G6PD gene specifically using SEQ ID NOs: 27 to 28, specifically amplify the HLA-DRB1 gene using SEQ ID NOs: 29 to 30, and HLA using SEQ ID NOs: 31 to 32 -Specifically amplify the DQA1 gene, specifically amplify the amplification quality control gene HGF using SEQ ID NOs: 33 to 34, specifically amplify the TPMT gene using SEQ ID NOs: 35 to 38, and SEQ ID NO: 39 Using to 40 to specifically amplify the NUDT15 gene, to specifically amplify the DPYD gene using SEQ ID NOs: 41 to 44, and to specifically amplify the UGT1A1 gene using SEQ ID NOs: 45 to 46, and SEQ ID NO: Using 47 to 50, the SLCO1B1 gene was specifically amplified, and the ABCG2 gene was specifically amplified using SEQ ID NOs: 51 to 52, and the SLC14A2 gene was specifically amplified using SEQ ID NOs: 53 to 56, and the sequence Specifically amplifying the C11orf65 gene using numbers 57 to 58, specifically amplifying the ACE gene using SEQ ID NOs: 59 to 60, and specifically amplifying the COQ2 gene using SEQ ID NOs: 61 to 62, Amplify the KCNJ11 gene specifically using SEQ ID NOs: 63 to 64, specifically amplify the NEDD4L gene using SEQ ID NOs: 65 to 66, and specifically amplify the ADRA1A gene using SEQ ID NOs: 67 to 68. And specifically amplify the AGTR1 gene using SEQ ID NOs: 69 to 70, specifically amplify the GNB3 gene using SEQ ID NOs: 71 to 72, and specific MMP3 gene using SEQ ID NOs: 73 to 74 It is amplified separately, and the PTGS gene can be specifically amplified using SEQ ID NOs: 75 to 76.

In the present invention, in the step (c), the snapshot analysis may be performed using one or more primers selected from the group consisting of primers of SEQ ID NOs: 77 to 124.

In the present invention, in the step (c), genotyping is, ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1, *2, *3, *17, CYP2C9 *1 ,*2,*3,*13,*14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52, *60,*XN (duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs5219, MMP3 rs3025058, NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*1B,*5,*15, TPMT *1,*3*6 It can be carried out by identifying one or more mutations selected from the group consisting of 1,*28 and VKORC1 rs9923231.

ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1, *2, *3, *17, CYP2C9 *1, *2, related to drug hypersensitivity reactions and drug side effects according to the present invention *3,*13,*14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,* XN (Duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T , KCNJ11 rs5219, MMP3 rs3025058, NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*1B,*5,*15, TPMT *1,*3C,*6*28, UGT , VKORC1 rs9923231 mutation detection kit and diagnostic method using it show high accuracy, especially for Asians, including Koreans, and can be quickly and easily diagnosed, preventing adverse drug reactions in advance and treatment effects. And it can be usefully used to increase the success rate.

1 shows the results of detecting 50 mutations for 27 genes related to drug reactions and drug side effects, including drug effects, using the kit according to the present invention.
2 is a table showing information on genes and their mutations used in the present invention.
3 is a table showing drug reactions and side effects that can be predicted by the method according to the present invention, and target genes therefor.
Figure 4 shows the sequence of the primer for amplification of the gene according to the present invention and information about it.
5 shows PCR conditions for amplifying a gene using the primer in the present invention.
Figure 6 shows the primer sequence and information for the snapshot analysis according to the present invention.
7 shows Exo-SAP IT treatment conditions for removing primers and dNTPs in amplified amplified products for the snapshot analysis in the present invention.
8 shows a snapshot condition according to the present invention.
9 shows the treatment conditions of Shrimp Alkaline Phosphatase (SAP) for the snapshot product in the snapshot analysis in the present invention.
10 is a schematic diagram showing an example of identifying a mutation and presenting a drug administration guide using the kit according to the present invention.

Definitions of terms used in the present invention are as follows.

'Genetic polymorphism' refers to a case in which genetic variation occurs with a frequency of at least 1% or more in a population. The insertion, deletion, or substitution of a single nucleotide in DNA is called single nucleotide polymorphism (SNP).

The term'polymorphism' refers to the arrangement in a sequence of genes that change within a population. Polymorphisms are made up of different “alleles”. This polymorphic arrangement can be identified by its position in the gene and the different amino acids or bases found therein. These amino acid variations are the result of two different alleles, two possible variant bases, C and T. Because the genotype is made up of two different distinct alleles, any of several possible variants can be observed in any one individual (eg, CC, CT or TT in this example). Individual polymorphisms are also known to those of skill in the art and are used, for example, in the Single Nucleotide Polymorphism Database (dbSNP) of Nucleotide Sequence Variation available on the NCBI website. It is an identifier (“reference SNP”, “refSNP” or “rs#).

The term “genotype” refers to a specific allele of a specific gene in a cell or tissue sample. 'Phenotype' refers to the morphological, physiological, or biochemical characteristics of an individual determined by the genotype. The genotype is distinct from the phenotype, and refers to the constitution of an individual's genes, and in a narrower sense, it refers to alleles that exist on one gene.

“Allele” or “allele” refers to one of two or more alternative forms of a gene that occupy the same chromosomal locus. The equivalent genetic marker of an allele is a genetic marker associated with the allele of interest, ie, it indicates a linkage disequilibrium with the allele of interest.

'Allele frequency' refers to the frequency (ratio or percentage) that an allele exists within an individual, within a lineage, or within a population of lineages. It can be estimated by averaging the allele frequency within a line or population or in a sample of individuals from a line or population.

'Risk' indicates that the incidence of the disease or condition is statistically high in the individual carrying the specific polymorphic allele compared to the incidence of the disease or condition in a member of the individual who does not carry the specific polymorphic allele.

"Nucleic acid" refers to a polynucleotide or oligonucleotide such as deoxyribonucleic acid (DNA), and if appropriate ribonucleic acid (RNA). The term also applies equally to analogs of either RNA or DNA prepared from nucleotide analogs (e.g., peptide nucleic acids), and single (sense or antisense) and double-stranded polynucleotides, as applied to the described embodiments. Include. In the present invention, the term "nucleic acid" and "nucleotide" are used in combination.

'Primer' is an oligonucleotide sequence that hybridizes to a complementary RNA or DNA target polynucleotide and serves as a starting point for the stepwise synthesis of polynucleotides from mononucleotides, for example by the action of nucleotidyl transferases occurring in polymerase chain reactions. Means.

The term “functional equivalent” refers to, for example, one or more substitutions, deletions or additions from a reference sequence, a net effect that does not result in various functional dissimilarities between the reference sequence and the subject sequence. It refers to both the nucleotide and nucleic acid sequences of the mutant sequence. Preferably, the nucleotide sequence has at least about 65% identity, more preferably at least about 75% identity, and most preferably about 95% identity. For the purposes of the present invention, sequences having substantially equivalent biological activity and substantially equivalent comparability characteristics are treated as substantial equivalents.

'Subject' or'patient' means any single individual in need of treatment, including humans, cattle, dogs, guinea pigs, rabbits, chickens, insects, and the like. In addition, any subject who participated in a clinical study trial showing no clinical manifestation of any disease, or a subject who participated in an epidemiological study or a subject used as a control group is included. In one embodiment of the present invention, humans were targeted.

“Tissue or cell sample” means a collection of similar cells obtained from tissues of a subject or patient. The source of the tissue or cell sample may be a fresh, frozen and/or preserved organ or tissue sample or solid tissue from a biopsy or aspirate; Blood or any blood component; It may be a cell at any point in the subject's pregnancy or development. The tissue sample can also be a primary or cultured cell or cell line.

All technical terms used in the present invention, unless otherwise defined, are used in the same sense as those of ordinary skill in the art generally understand in the related field of the present invention. In addition, although preferred methods or samples are described in the present specification, those similar or equivalent are included in the scope of the present invention. The contents of all publications referred to herein by reference are incorporated into the present invention.

Hereinafter, the present invention will be described in detail.

The present invention is based on the fact that various drug reactions or adverse drug reactions (ADRs) are associated with drug metabolism, transport, and pharmacodynamics total 27 genes, 50 mutations, ABCG2, ACE, ADRA1A, AGTR1, C11orf65, COQ2, CYP2C19 , CYP2C9, CYP2D6, CYP4F2, DPYD, G6PD, GNB3, HLA-B, HLA-DQA1, HLA-DRB1, Internal Control HGF, KCNJ11, MMP3, NEDD4L, NUDT15, PTGS2, SLC14A2, SLCO1A1, TPMT, and UGT genes Subtype, that is, a kit for detecting the presence of a specific mutation in each gene and a specific mutation using the kit to quickly and accurately detect the drug side effects for cancer and chronic disease-related multi-prescription drugs and for personalized drug treatment Provides a method for diagnosing multiple high-speed drug genes.

Specifically, the kit and method of the present invention are ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1, *2, *3, *17, CYP2C9 *1, *2, *3 ,*13,*14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN ( Duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs5219, MMP3 rs3025058, NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*1B,*5,*15, TPMT *1,*3C,*1,*28 and VKOVRC,*1,*28 and VKO1A1 It is characterized by analyzing the presence of 50 mutations in a total of 27 genes of rs9923231 (see FIG. 2).

Hereinafter, the configuration and features of the kit and method according to the present invention will be described for example as follows, but are not limited thereto.

Multi-speed drug gene diagnosis kit for predicting drug side effects and personalized drug treatment for cancer and chronic diseases related multi-prescription drugs

The multi-high-speed pharmacokinetics diagnostic kit for predicting drug side effects and personalized drug treatment for cancer and chronic disease-related multi-prescription drugs of the present invention includes primers of SEQ ID NOs: 1 to 124 (see FIGS. 4 and 6), and ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1,*2,*3,*17, CYP2C9 *1,*2,*3,*13,*14, CYP2D6 *1 *2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN (duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs5219, MMP3 rs304141,116,232,DD4 PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A, *1B, *5, *15, TPMT *1, *3C, *6, UGT1A1 *1, *28 and at least one mutation selected from the group consisting of VKORC1 rs9923231 It is characterized by determining the presence or absence (see Fig. 2).

Specifically, the kit includes 2X PCR Amplification Mix, 2X Multiplex PCR Amplification Mix, amplification primer mix 1 (APM1), amplification primer mix 2 (APM2), amplification primer mix 3 (APM3), amplification primer mix 4 (APM4), amplification Primer Mix 5 (APM5), Amplification Primer Mix 6 (APM6), Amplification Primer Mix 7 (APM7), Amplification Primer Mix 8 (APM8), Amplification Primer Mix 9 (APM9), Amplification Primer Mix 10 (APM10), Snapshot Multi Flex Reagent (SNaPshot MR), Snapshot Primer Mix 1 (SPM1), Snapshot Primer Mix 2 (SPM2), Snapshot Primer Mix 3 (SPM3), Snapshot Primer Mix 4 (SPM4), Snapshot Primer Mix 5 (SPM5) ), control DNA and nuclease-free water.

At this time, for example, 2X PCR Amplification Mix is sold by Solgent, and per 10 ul of 10X EF tag polymerase buffer 2 ul, 5X Band Doctor 1 ul, 2.5mM dNTP 2 ul, 2.5U EF tag polymerase 0.25 ul and nuclease-free water 4.75 ul, 2X Multiplex PCR Amplification Mix is sold by Solgent, 10X H tag polymerase buffer per 10 ul per time 2 ul, 5X Band Doctor 1 ul, 2.5mM dNTP 2 ul, 2.5U EF tag polymerase 0.25 ul and nuclease free water 4.75 ul.

In addition, the kit may further include Shrimp Alkaline Phosphatase (SAP) and/or Exonuclease 1 (Exo).

As shown in Figure 4, the amplification primer mix 1 is composed of the primers of SEQ ID NO: 1 to SEQ ID NO: 4, the amplification primer mix 2 is composed of the primers of SEQ ID NOs: 5 to 6, and the amplification primer mix 3 is sequence Consists of primers of SEQ ID NOs: 7 to 8, amplification primer mix 4 is composed of primers of SEQ ID NOs: 9 to 24, amplification primer mix 5 is composed of primers of SEQ ID NOs: 25 to 28, amplification primer mix 6 is sequence Consists of primers of Nos. 29 to 44, amplification primer mix 7 consists of primers of SEQ ID NOs: 45 to 46, amplification primer mix 8 is composed of primers of SEQ ID NOs: 47 to 56, and amplification primer mix 9 is of SEQ ID NO: 57 It consists of the primers of to 66, and amplification primer mix 10 consists of the primers of SEQ ID NOs: 67 to 76. The amplification primer mix (APM) 4, 6, 7, 8, 9 and 10 were amplified using 2X Multiplex PCR Amplification Mix, and the amplification primer mix (APM) 1, 2, 3 and 5 used 2X PCR Amplification Mix. To amplify.

The snapshot multiplex reagent uses a commercially available reagent from Applied biosystems, and as shown in FIG. 6, the snapshot primer mix (SPM) 1 consists of primers of SEQ ID NOs: 77 to 87, and the snapshot Primer mix 2 consists of primers of SEQ ID NOs: 88 to 99, snapshot primer mix 3 consists of primers of SEQ ID NOs: 100 to 109, and snapshot primer mix 4 consists of primers of SEQ ID NOs: 110 to 119, snap Shot primer mix 5 consists of primers of SEQ ID NOs: 120 to 124. Nuclease-free water uses a commercially available reagent from Solgent.

The kit of the present invention uses genomic DNA prepared from biological samples such as human or mammalian blood, whole blood nucleic acids, etc., using a single base extension (SBE, SNaPshot, Mutation identification by primer extension) for a total of 50 mutations of 27 genes. ), and is characterized in that it is a kit that helps predict related drug reactions and drug side effects (ADR).

As an embodiment of the present invention, nucleotide 8825 of the ABCG2 gene (NCBI Reference Sequence: NC_000004.10) is converted from C to A (rs2231142) using the kit, and the ACE gene (NCBI Reference Sequence: NC_000017.10). TTTTTTTTTTTGAGACGGAGTCTCGCTCTGTCGCCCATACAGTCACTTTT insertion or deletion of nucleotides 11435-11436 (rs1799752), nucleotide 94459 of ADRA1A gene (NCBI Reference Sequence: NC_000008.11) is T to C conversion (rs1048101), AGTR1 gene (NCBI Reference Sequence: NC_000003.12) is converted from A to C (rs5186), the 49126th nucleotide of the C11orf65 gene (NCBI Reference Sequence: NC_000011.10) is converted from G to T (rs11212617), and the COQ2 gene (NCBI Reference Sequence: NC_000004.12) is converted from C to G (rs4693075), the 470865 nucleotide of the DPYD gene (NCBI Reference Sequence: NC_000001.11) is converted from G to A (rs3918290), from A to 838532 nucleotide Conversion to T (rs67376798), nucleotide 11716 of the G6PD gene (NCBI Reference Sequence: NC_000023.10) is converted from A to T (rs5030872), nucleotide 11737 is converted from C to T (rs5030868), GNB3 gene (NCBI Reference Sequence: NC_000012.12), nucleotide 4424 is converted from C to T (rs5443), KCNJ 11 gene (NCBI Refe rence Sequence: NC_000011.9) is converted from A to G (rs5219), the -1676_-1675 nucleotide of the MMP3 gene (NCBI Reference Sequence: NC_000011.10) is the T insertion (rs3025058), the NEDD4L gene ( NCBI Reference Sequence: NC_000018.10) nucleotide 99899 is converted from G to A (rs4149601), NUDT15 gene (NCBI Reference Sequence: NC_000013.11) has 7973 nucleotide converted from C to T (rs116855232), PTGS2 gene ( NCBI Reference Sequence: NC_000001.11) at nucleotide -899 is G to C conversion (rs20417), SLC14A2 gene (NCBI Reference Sequence: NC_000018.10) at nucleotide 48254 is G to A conversion (rs1123617), 57730 The nucleotide is used to detect the G-to-A conversion (rs3745009) and the G-to-A conversion (rs9923231) of nucleotide-1639 of the VKORC1 gene (NCBI Reference Sequence: AY587020).

In addition, the CYP2C19 gene (NCBI Reference Sequence: NG_008384.2, The Human Cytochrome P450 (CYP) Allel Nomenclature reference) *1 (Wild type), *2 (19154th nucleotide converted from G to A, rs4244285), *3 ( 17948th nucleotide is converted from G to A, rs4986893), *17 (-806th nucleotide is converted from C to T, rs12248560), CYP2C9 gene (NCBI Reference Sequence: NG_008385.1, The Human Cytochrome P450 (CYP) Allel Nomenclature Reference) of *1 (Wild type), *2 (conversion from nucleotide C to T at 3608, rs1799853), *3 (conversion from nucleotide at 42614 A to C, rs1057910), *13 (T to C at nucleotide 3276 Conversion to, rs72558187), *14 (transformation of nucleotide 3552 from G to A, rs72558189), *1 (wild type) of the CYP2D6 gene (NCBI Reference Sequence: M33388, The Human Cytochrome P450 (CYP) See Allel Nomenclature), *10B (100th nucleotide C to T conversion, rs1065852), *49 (1611th nucleotide T to A conversion, rs1135822), *14B (1758th nucleotide G to A conversion, rs5030865), *4 ( G to A conversion of nucleotide 1846, rs3892097), *60 (TA insertion of nucleotide 1887), *21 (C insertion of nucleotide 2573-2574, rs72549352), *2 (C to T conversion of nucleotide 2850 , rs16947), *41 (transformation of nucleotide 2988 from G to A, rs28371725), *52 (transformation of nucleotide 3877 from G to A, rs28371733 ), *18 (GTGCCCACT duplication of nucleotide 4125-4133), *5 (CYP2D6 deletion), *XN (CYP2D6 duplication), CYP4F2 gene (NCBI Reference Sequence: AF467894.1, The Human Cytochrome P450 (CYP) Allel Nomenclature reference) *1 (wild type), *3 (conversion of nucleotide 1297 from G to A, rs2108622), SLCO1B1 gene (NCBI Reference Sequence: NG_011745.1, SLCO Nomenclature (refer to SLCO Nomenclature) *1A (wild type), *1B (50611 nucleotide A to G conversion, rs2306283), *5 (52422 nucleotide T to C conversion, rs4149056), *15 (50611 nucleotide A to G conversion of, rs2306283 and T to C conversion of nucleotides 52422, rs4149056), TPMT gene (NCBI Reference Sequence; NM_000367.2, refer to TPMT Nomenclature) *1 (Wild type), *3C (A to G conversion of the 719th nucleotide, rs1142345), *6 (A to T conversion of the 539th nucleotide, rs75543815), UGT1A1 gene *1 (Wild type) of (NCBI Reference Sequence: NT_005120.25, refer to UGT1A1 Nomenclature), *28 (TA deletion and insertion of nucleotide -39, rs3064744), HLA-B gene (NCBI Reference Sequence: NG_023187.1) *57 (conversion of nucleotide 384 from C to G, conversion of nucleotide 667 from G to A), *58 (conversion of nucleotide 384 from C to G), HLA-DQA1 gene (NCBI Reference Sequence: NG_032876. It is used to detect *02 of 1) (conversion of nucleotide 91 from A to G) and *07 (conversion of nucleotide 8240 from G to A) of HLA-DRB1 gene (NCBI Reference Sequence: NG_029921.1). To identify the *07 allele of the HLA-DRB1 gene, nucleotide 473 of the HGF gene (NCBI Reference Sequence: NC_000017.11) can be used as an amplification quality control gene.

In addition, in the present invention, the drug is voriconazole, piroxicam, aspirin, celecoxib, diclofenac, tramadol, ibuprofen, omeprazole, esomeprazole, lansoprazole, pantoprazole, rabeprazole, simbassatin, seri Vastatin, pravastatin, rosuvastatin, atorvastatin, clopidogrel, flecainide, propaphenone, nifedipine, captopril, quinacryl, enalapril, lisinopril, telmisartan, losartan, metoprolol, irbesartan , Candesartan, carvedirol, hydrochlorothiazide, warfarin, glimepiride, glibenclamide, gliclazide, metformin, allopurinol, lapatinib, tamoxifen, cisplatin, mercaptopurine, pazopanib, capecitabine , Fluorouracil, irinotecan, azathioprine, rasburicase, and similar drugs, and the drug side effects are Stevens-Johnson syndrome, toxic epidermal necrosis dissolution, irritability syndrome, severe liver injury, muscle It may be selected from the group consisting of diseases.

In addition, the kit according to the present invention is ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1, *2, *3, *17, CYP2C9 *1, *2, *3, * 13,*14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN (duplicate) , CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 47319T, KCNJ11 rs5219T, KCNJ11 rs5219 MMP3 rs3025058, NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*1B,*5,*15, TPMT *1,*3C,*6, UGT1A1 *1,*23231 And probes for detecting one or more mutant alleles in the group consisting of.

As used herein, the probe means "any substance useful for detecting other substances". Thus, the probe may be an oligonucleotide or a conjugated oligonucleotide that specifically hybridizes to a specific region within the allele of interest. Conjugated oligonucleotide refers to an oligonucleotide that covalently binds to a chromophore or ligand containing molecule (eg, antigen), which is highly specific to a receptor molecule (eg, antigen specific antibody). The probe may be a PCR primer for amplifying a specific region within the allele of interest together with other primers. Furthermore, the probe may be an antibody that recognizes an allele of interest or a protein product of the allele.

The kit may further include tools and/or reagents for collecting a biological sample from a patient, as well as tools and/or reagents for preparing genomic DNA, RNA, cDNA or protein from the sample. For example, it may include PCR primers for amplifying the relevant region of genomic DNA. The kit may contain probes for genetic factors useful for pharmacogenomic profiling. In addition, in the use of such a kit, a labeled oligonucleotide can be used to easily identify it during analysis. Examples of labels that can be used include radioactive labels, enzymes, fluorescent compounds, streptavidin, avidin, biotin, magnetic moieties, metal binding moieties, antigen or antibody moieties, and the like.

In particular, in the present invention, ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1, *2, *3, *17, CYP2C9 *1, *2, *3, *13, * 14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN (duplicate), CYP4F2 * 1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs5219, MMP3 NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*1B,*5,*15, TPMT *1,*3C,*6, UGT1A1 *1,*28, VKORC1 rs9923231 It is preferred to include a set of primers for amplification of one or more mutations and mutant regions, for example, primer sets represented by SEQ ID NOs: 1-76.

Mutation analysis method using the kit according to the present invention

The present invention uses the kit as a total of 27 genes, 50 mutations related to drug reactions and drug side effects, ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1,*2,* 3,*17, CYP2C9 *1,*2,*3,*13,*14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,* 41,*49,*52,*60,*XN (duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1* 02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs5219, MMP3 rs3025058, NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*1B,*1A,*1B,* *3C, *6, UGT1A1 *1, *28 and VKORC1 rs9923231 provides a method to quickly and accurately detect and analyze mutations in each gene by detecting the presence of mutations, and drug reactions including drug effects based on the identified mutations , Drug side effects assessment, drug identification, etc.

The allele can be detected, for example, by directly detecting the region/nucleotide in the allele using genomic DNA prepared from a biological sample such as blood, saliva, urine, or hair. Alleles can also be detected, for example, by serological methods or microcytotoxic methods.

It can also be determined by detection of an equivalent genetic marker of the allele, and the equivalent genetic marker is, for example, a single nucleotide polymorphism (SNP), a microsatellite marker, or other types of It is polymorphic. In the present invention, the use of SNP is preferred. The equivalent genetic marker of an allele is a genetic marker associated with the allele of interest, ie, it indicates a linkage disequilibrium with the allele of interest.

ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1,*2,*3,*17, CYP2C9 *1,*2,*3,*13,*14, CYP2D6 ,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN (duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs5219, MMP3 rs30L15 rs5219, MMP3 rs30L rs5219, MMP3 rs30L15 rs5219, MMP3 rs30L , PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A, *1B, *5, *15, TPMT *1, *3C, *6, UGT1A1 *1, *28 and VKORC1 rs9923231 in total 27 genes, 50 mutations The presence of a genetic marker such as the back can be determined by direct detection of a label or a specific region therein.

For example, SNP analysis may be performed using a real time PCR system, or may be performed through direct determination of the nucleotide sequence of a nucleic acid by a dideoxy method, or a probe containing a sequence of a SNP site or A probe complementary thereto is hybridized with the DNA and the nucleotide sequence of the polymorphic site is determined by measuring the degree of hybridization obtained therefrom, or the allele-specific probe hybridization method, the allele-specific amplification method (allele- specific amplification), sequencing, 5'nuclease digestion, molecular becon assay, oligonucleotide ligation assay, size analysis And single-strand conformation polymorphism.

DNA products obtained from PCR can be detected using sequence-specific probes such as hydrolysis probes (Taqman, Becons, Scorpions) and matching probes, for example. These probes are designed to bind to the area of interest. The PCR product can also be detected by a DNA-binding agent.

The presence of an allele of interest can also be determined by detection of an equivalent genetic marker of the allele. Genetic markers close to the allele of interest tend to co-segregate with the allele or exhibit linkage disequilibrium. Thus, the presence of these markers (equivalent genetic markers) is an indicator of the presence of the allele of interest and thus the risk of developing ADR.

Useful equivalent genetic markers are usually about 200 kb or less (eg, 100 kb, 80 kb, 60 kb, 40 kb or 20 kb) from the allele of interest. The presence or absence of an equivalent genetic marker can be detected using the above-described method or a method known in the art.

In addition, RNA, cDNA, or protein products of an allele of interest can be detected to determine the presence or absence of the allele.

As such, the presence of mutations in the gene of interest can be detected by a sequence-based genotyping method (Sanger sequencing) or allele-specific PCR in various known methods, but the sequencing method takes a lot of time to read and a special program for reading. And the possibility of error due to allele-specific PCR. However, specific mutations to be detected in the present invention ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1, *2, *3, *17, CYP2C9 *1, *2, *3 ,*13,*14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN ( Duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs5219, MMP3 rs3025058, NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*1B,*5,*15, TPMT *1,*3C,*1,*28 and VKOVRC,*1,*28 and VKO1A1 Analyzing rs9923231, it is a method of reducing sequencing, so it is efficient in reducing accuracy and time.

In addition, the method of the present invention can be used to analyze an effective genome-side association structure by identifying the structure of the haplotype within the gene and the SNP representing each haplotype.

Thus, the method of the present invention uses a method of detecting 50 mutations (single base polymorphism, SNP) of a total of 27 genes, and includes the following steps:

(a) ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1,*2,*3,*17, CYP2C9 *1,*2,*3,*13,*14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN (duplicate), CYP4F2 *1, *3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs5241458, MMP4DDL rs3025019601 , NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A, *1B, *5, *15, TPMT *1, *3C, *6, UGT1A1 *1, *28, VKORC1 rs9923231 Determining a detectable SNP (see FIG. 2);

(b) ABCG2, ACE, ADRA1A, AGTR1, C11orf65, COQ2, CYP2C19, CYP2C9, CYP2D6, CYP4F2, DPYD, G6PDGNB3, HLA-B, HLA-DQA1, HLA-DRB1, Internal Control HGF, NEDD15, NUDT15, PTGS2, NUDT15 , SLC14A2, SLCO1B1, TPMT, UGT1A1, performing a multiple polymerase chain reaction to amplify VKORC1; And

(c) performing a snapshot (SNaPshot) analysis using multiple single base primers to analyze the SNP for the product to confirm the genotype.

At this time, the multiple polymerase chain reaction in step (b) may be performed using one or more primers selected from the group consisting of primers of SEQ ID NOs: 1 to 76.

Specifically, CYP2D6 was specifically amplified using SEQ ID NOs: 1 to 8, CYP2C19 was specifically amplified using SEQ ID NOs: 9 to 14, and CYP4F2 was specifically amplified using SEQ ID NOs: 15 to 16. And, using SEQ ID NOs: 17 to 22, the CYP2C9 gene is specifically amplified, and the VKORC1 gene is specifically amplified using SEQ IDs: 23 to 24, and the HLA-B gene is specific using SEQ ID NOs: 25 to 26. Amplify specifically, amplify the G6PD gene specifically using SEQ ID NOs: 27 to 28, specifically amplify the HLA-DRB1 gene using SEQ ID NOs: 29 to 30, and HLA using SEQ ID NOs: 31 to 32 -Specifically amplify the DQA1 gene, specifically amplify the amplification quality control gene HGF using SEQ ID NOs: 33 to 34, specifically amplify the TPMT gene using SEQ ID NOs: 35 to 38, and SEQ ID NO: 39 Using to 40 to specifically amplify the NUDT15 gene, to specifically amplify the DPYD gene using SEQ ID NOs: 41 to 44, and to specifically amplify the UGT1A1 gene using SEQ ID NOs: 45 to 46, and SEQ ID NO: Using 47 to 50, the SLCO1B1 gene was specifically amplified, and the ABCG2 gene was specifically amplified using SEQ ID NOs: 51 to 52, and the SLC14A2 gene was specifically amplified using SEQ ID NOs: 53 to 56, and the sequence Specifically amplifying the C11orf65 gene using numbers 57 to 58, specifically amplifying the ACE gene using SEQ ID NOs: 59 to 60, and specifically amplifying the COQ2 gene using SEQ ID NOs: 61 to 62, Amplify the KCNJ11 gene specifically using SEQ ID NOs: 63 to 64, specifically amplify the NEDD4L gene using SEQ ID NOs: 65 to 66, and specifically amplify the ADRA1A gene using SEQ ID NOs: 67 to 68. And specifically amplify the AGTR1 gene using SEQ ID NOs: 69 to 70, specifically amplify the GNB3 gene using SEQ ID NOs: 71 to 72, and specific MMP3 gene using SEQ ID NOs: 73 to 74 It is amplified separately, and the PTGS gene can be specifically amplified using SEQ ID NOs: 75 to 76.

The multiple single base primers for the snapshot analysis in step (c) use any one primer selected from the group consisting of SEQ ID NOs: 77 to 124.

Genotype confirmation in step (c) is ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1, *2, *3, *17, CYP2C9 *1, *2, *3 ,*13,*14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN ( Duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs5219, MMP3 rs3025058, NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*1B,*5,*15, TPMT *1,*3C,*1,*28 and VKOVRC,*1,*28 and VKO1A1 It may be to identify one or more mutations selected from the group consisting of rs9923231.

As an embodiment, the method may be performed in the following manner.

(a) Target ABCG2, ACE, ADRA1A, AGTR1, C11orf65, COQ2, CYP2C19, CYP2C9, CYP2D6, CYP4F2, DPYD, G6PDGNB3, HLA-B, HLA-DQA1, HLA-DRB1, Internal Control HGF, NE4DDL, NUDT15, KCNJ11M , PTGS2, SLC14A2, SLCO1B1, TPMT, UGT1A1 and specific sites of VKORC1.

In an embodiment of the present invention, SNPs and mutation sites were included to be amplified, and exons 2 to 4 of HLA-B, exon 1 of HLA-DQA1, exon 2 of HLA-DRB1, and exon 2 of HGF Was allowed to amplify. At this time, an appropriate known method may be selected and used, for example, a method such as multiplex polymerase chain reaction (multiplex PCR) may be used.

In the examples of the present invention, primer sets of SEQ ID NOs: 1 to 76 were used for the amplification, and their sequences and uses are also included in the scope of the present invention. In addition, as shown in Figure 4, each primer pair is specific to a specific gene.

(c) SNP is analyzed by performing a snapshot analysis method through multiple single base primer expansion.

The primers to be used in this step are ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1,*2,*3,*17, CYP2C9 *1,*2,*3,*13, *14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN (duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs3025058, MMP3 , NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A, *1B, *5, *15, TPMT *1, *3C, *6, UGT1A1 *1, *28 and each mutation of VKORC1 rs9923231 Is designed to be aligned in the front of which can be distinguished, and expansion reaction is performed using the above primers at multiple single base polymorphism (SNP) sites.

Multiple single base primers used in an embodiment of the present invention are SEQ ID NOs: 77 to 124, and their sequences and uses are also included in the scope of the present invention. In addition, as shown in Figure 6, each primer pair is specific to a specific gene.

Thereafter, by analyzing the peak of single nucleotide polymorphism (SNP) for the product to be produced to confirm the genotype, ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1,*2,*3, *17, CYP2C9 *1,*2,*3,*13,*14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41, *49,*52,*60,*XN (duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs5219, MMP3 rs3025058, NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*15, TP *1,*1B,*5 , *6, UGT1A1 *1, *28 and VKORC1 rs9923231 It is determined whether or not at least one mutation selected from the group consisting of (see Fig. 1).

Thus, detection of such mutations by any available method can be used for diagnostic purposes such as response of drugs, early detection of susceptibility to drug side effects, prognosis, diagnosis and treatment for patients.

Another aspect of the present invention by the above method is a method of pharmacogenomic profiling. “Pharmacogenomic profiling” refers to the determination of genetic factors present in a subject, related to a disease or medical condition, particularly adverse reactions to a drug. This method is ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1,*2,*3,*17, CYP2C9 *1,*2,*3,*13,*14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN (duplicate), CYP4F2 *1, *3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs5241458, MMP4DDL rs3025019601 , NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A, *1B, *5, *15, TPMT *1, *3C, *6, at least selected from the group consisting of UGT1A1 *1, *28 and VKORC1 rs9923231 Determining the presence of one mutation. The method may include determination of other genetic factors as needed. Such other genetic factors may be associated with the predisposition of any disease or medical condition, including ADR.

Drug response and drug side effects (ADR) evaluation using each mutation detection result

In yet another aspect, the present invention relates to a method for assessing the risk of causing adverse drug reactions in a patient in response to a drug, comprising a method of determining the genotype using a biological sample from a patient by the above method.

That is, the genetic label, ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1, *2, *3, *17, CYP2C9 *1, *2, *3, *13, *14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN (duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs3025058, MMP3 , NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A, *1B, *5, *15, TPMT *1, *3C, *6, UGT1A1 *1, *28 and VKORC1 rs9923231 exist or A method of assessing a patient's risk of developing a drug response or adverse drug reactions (eg, Stevens-Johnson syndrome, toxic epidermal necrolysis, or irritability syndrome, etc.) based on the absence is provided. The presence of the allele is an indicator of the risk of adverse drug reactions.

The drugs are preferably voriconazole, piroxicam, aspirin, celecoxib, diclofenac, tramadol, ibuprofen, omeprazole, esomeprazole, lansoprazole, pantoprazole, rabeprazole, simbassatin, cerivastatin, pravastatin , Rosuvastatin, atorvastatin, clopidogrel, flecainide, propaphenone, nifedipine, captopril, quinacryl, enalapril, lisinopril, telmisartan, losartan, metoprolol, irbesartan, candesartan , Carvedirol, hydrochlorothiazide, warfarin, glimepiride, glibenclamide, gliclazide, metformin, allopurinol, lapatinib, tamoxifen, cisplatin, mercaptopurine, pazopanib, capecitabine, fluorouracil , Irinotecan, azathioprine, rasburicase, and similar drugs.

In the present invention, in the drug compound, the drug compound itself or at least one hydrogen in the drug is halo, hydroxy, acylamino, alkyl, alkenyl, alkynyl, alkoxy, aryloxy, aryl, aryloxyaryl, carboxy , Carboxyalkyl, carboxy-substituted alkyl, carboxy-cycloalkyl, carboxy substituted cycloalkyl, carboxyaryl, carboxy-substituted aryl, carboxyheteroaryl, carboxy-substituted heteroaryl, carboxyheterocyclic, carboxy-substituted Heterocyclic, cycloalkyl, substituted alkyl, substituted alkoxy, substituted aryl, substituted aryloxy, substituted aryloxyaryl, substituted cycloalkyl, heteroaryl, substituted heteroaryl, heterocyclic or substituted hetero It refers to a compound such as the drug, except that it is substituted with a cyclic group. The number of carbon atoms contained in the substituent may be 0 to 10, 0 to 6, 0 to 4, or 0 to 2.

For these drugs, if the probability of developing ADR is higher than that of the general population, the patient is at "risk" of ADR.

The patient's probability of inventing ADR is at least about 1.5 times, more preferably at least about 2 times, even more preferably at least about 3, 4, 5, 6, 7, 8 or 9 times the probability of developing ADR in the general population, And most preferably at least about 10 times. The probability may be determined using any method known in the art, such as using the incidence of risk factors.

The “risk” factors of ADR are factors related to ADR. The sensitivity of the risk factor is preferably at least about 40%, more preferably at least about 50%, 60%, 70%, 80%, 85% or 90%. Most preferably, the sensitivity is at least 95%. The “sensitivity” of a risk factor for predicting ADR is the percentage of ADR patients with that risk factor. For example, if all SJS patients have allele A, the sensitivity of allele A to predict SJS is 100%. If 20 of 40 SJS patients have allele B, then the sensitivity of allele B to predict SJS at that time is 50%.

ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1,*2,*3,*17, CYP2C9, associated with drug reactions and adverse drug reactions (ADR) with a sensitivity of at least about 40% *1,*2,*3,*13,*14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,* 52,*60,*XN (duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1* 07, Internal Control HGF 473T, KCNJ11 rs5219, MMP3 rs3025058, NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*1B,*5,*15,*6,*6, TP UGT1A1 *1, *28 and VKORC1 rs9923231 mutations can be used as risk factors in the present invention. Preferably, the sensitivity of the risk factor is at least about 50%, 60%, 70%, 80%, 85% or 90%. More preferably, the sensitivity is at least 95%.

The present invention also relates to drug reactions and adverse drug reactions (ADR) ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1, *2, *3, *17, CYP2C9 *1, *2,*3,*13,*14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,* 60,*XN (duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs5219, MMP3 rs3025058, NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*1B,*5,*15, TPMT *1,*6 ,*28, VKORC1 This includes methods of identifying similar compounds that cause ADR (eg, SJS/TEN, HSS, severe liver injury, muscle disease, etc.) in patients carrying a mutation of rs9923231.

As described above, the present invention is ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1, *2, *3, *17, CYP2C9 *1, *2, *3, *13, *14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN (duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs3025058, MMP3 , NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*1B,*5,*15, TPMT *1,*3C,*6, UGT1A1 *1,*28, VKORC1 rs9923231, fast and accurate It covers all uses including detection methods and drug side effects assessment, drug identification, etc., in which these results can be utilized.

<Example>

Hereinafter, the present invention will be described in more detail through examples. These examples are for illustrative purposes only, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not construed as being limited by these examples.

Unless otherwise indicated, nucleic acids are written in a 5'->3' orientation from left to right. Numerical ranges recited within the specification include the numbers defining the range, and include each integer or any non-integer fraction within the defined range.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in practice to test the present invention, preferred materials and methods are described herein.

Example 1: multiplex polymerase chain reaction (multiplex polymerase chain reaction, multiplex PCR)

As shown in Fig. 4, the polymerase chain reaction was performed according to the conditions shown in Fig. 5 using an appropriate amplification primer mix according to the desired gene. Polymerase chain reaction was carried out using ABI 9700 (Applied Biosystems, Foster, USA).

For example, in the case of APM1, the reaction solution was configured by matching the final 20ul to include 100ng of the request sample, 10ul of 2X PCR Amplification Mix, and 4ul of amplification primer mix (APM1). After the reaction solution was treated at 94° C. for 5 minutes, it was reacted 35 times at 98° C. 20 seconds, 64° C. 30 seconds, and 72° C. 3 minutes each, and the final extension reaction was performed at 72° C. for 5 minutes.

Next, transfer the polymerase chain reaction product to an ice water bath, mix 2 ul of the PCR product and 1 ul of 6X loading buffer on a parafilm, and mix 100 bp and 1 kb on a 1% agarose gel. 2 ul of a DNA ladder was mixed with 0.5X TBE buffer, followed by electrophoresis at 100 V for 30 minutes.

Thereafter, the 1% agarose gel was stained with gel red containing a fluorescent substance to confirm the DNA size. To remove the primer and dNTP in the sample after checking the size of the PCR product, ExoSAP-IT (USB corp., Cleveland, USA) was treated according to the conditions shown in FIG. 7 and then reacted at 37°C for 35 minutes and 80°C for 15 minutes. Made it.

Example 2: Allele test using multiplex single based primer extension (SNaPshot assay)

2-1. Primer design by type

Primers of SEQ ID NOs: 77 to 124 were used as primers for each type for multiplex single based primer extension (SNaPshot assay) (see FIG. 6).

ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1,*2,*3,*17, CYP2C9 *1,*2,*3 ,*13,*14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN ( Duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs5219, MMP3 rs3025058, NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*1B,*5,*15, TPMT *1,*3C,*1,*28,KOGT1A1 It was designed to be aligned with the nucleotide immediately before the mutation that can distinguish rs9923231.

2-2. Snapshot method

For the snapshot analysis, a polymerase chain reaction product was used as a template, and as shown in FIG. 6, a multiple single base primer expansion reaction was performed according to the conditions shown in FIG. 8 using a snapshot primer mix containing a specific primer.

For example, in the case of Snapshot Primer Mix 1 (SPM1), the primers of SEQ ID NOs: 77 to 87 were used to extend multiple single base primers using 11 types of primers designed with different lengths for 11 single polymorphism sites. The reaction was carried out.

In the case of the multiple single base primer extension reaction, in the case of Snapshot Primer Mix 1 (SPM1), the snapshot multiplex reagent (SNaPshot MR) of the kit according to the present invention 1 ul, 1/2 term buffer 4 ul, Exo-SAP- It consisted of 7.5 ul of IT product and 1 ul of Snapshot Primer Mix 1 (SPM1).

The prepared products were reacted 40 times at 96°C for 10 seconds, 50°C for 5 seconds, and 60°C for 30 seconds, followed by adding 1.2 ul (1 U/ul) of SAP according to the conditions shown in FIG. 9 to 37°C for 60 minutes, The reaction was carried out at 65° C. for 15 minutes to remove the remaining ddNTP.

To 1 ul of the reaction product, 0.5 ul of GeneScan-120 LIZ size standard and 8.5 ul of Hi-Di formamide were added and analyzed using an ABI 3500 gene analyzer (Applied biosystems, Foster, USA).

2-3. Single base polymorphism (SNP) analysis

The extension reaction product was obtained by combining ddNTPs with complementary fluorescence attached to each single-base polymorphic position at the end of each primer 3′, and each peak for each SNP was analyzed. GeneMapper v5.0 (Applied Biosystems, Foster, USA) was used as the software, and according to the analysis result, ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1,*2,*3,* 17, CYP2C9 *1,*2,*3,*13,*14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,* 49,*52,*60,*XN (duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA -DRB1*07, Internal Control HGF 473T, KCNJ11 rs5219, MMP3 rs3025058, NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*1B,*5,*15 *6, UGT1A1 *1, *28, VKORC1 rs9923231 was confirmed to exist.

And the results obtained through the experiment were compared with the gene sequence information (sequencing data) obtained by the standard test method (gold standard, Sequencing).

ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1,*2,*3,*17, CYP2C9 *1,*2,*3,*13,*14, CYP2D6 ,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN (duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs5219, MMP3 rs30L15 rs5219, MMP3 rs30L rs5219, MMP3 rs30L15 rs5219, MMP3 rs30L , PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A, *1B, *5, *15, TPMT *1, *3C, *6, UGT1A1 *1, *28, VKORC1 rs9923231 at the same time test results are shown in FIG. Shown.

Through these results, using the kit of the present invention, drug reaction and drug side effects (ADR) related genes ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1, *2, *3 ,*17, CYP2C9 *1,*2,*3,*13,*14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41 ,*49,*52,*60,*XN (duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02 , HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs5219, MMP3 rs3025058, NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*15, TP *1A,*1B,*5 It can be seen that the presence of one or more mutations selected from the group consisting of 3C, *6, UGT1A1 *1, *28, and VKORC1 rs9923231 can be quickly and accurately confirmed.

Therefore, by using the present invention, the presence or absence of each of the specific mutations can be detected at a high speed, so that it can be effectively utilized in a method for predicting and diagnosing risk related to drug reactions and drug side effects.

So far, the present invention has been looked at around its preferred embodiments. Those of ordinary skill in the art to which the present invention pertains will be able to understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered from an illustrative point of view rather than a limiting point of view. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the scope equivalent thereto should be construed as being included in the present invention.

<110> SPMED Co.,Ltd. <120> Multiplex drug gene diagnosis kit for predicting drug side effects on chronic diseases and cancer related multi-prescription drugs and personalized drug treatment <130> DPP190142KR <160> 124 <170> KoPatentIn 3.0 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> CYP2D6 Amp100F <400> 1 ccatttggta gtgaggcagg t 21 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> CYP2D6 Amp1887R <400> 2 cctgcagaga ctcctcggtc t 21 <210> 3 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> CYP2D6 Amp2850F <400> 3 tggggcctga gacttgtcca gg 22 <210> 4 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> CYP2D6 AmpR <400> 4 actgagccct gggaggtagg ta 22 <210> 5 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> CYP2D6 DelF <400> 5 acctctctgg gccctcaggg a 21 <210> 6 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> CYP2D6 DelR <400> 6 caggcatgag ctaaggcacc cagac 25 <210> 7 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> CYP2D6 DupF <400> 7 cctcaccaca ggactggcca cc 22 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CYP2D6 DupR <400> 8 cacgtgcagg gcacctagat 20 <210> 9 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CYP2C19*2F <400> 9 aattacaacc agagcttggc 20 <210> 10 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> CYP2C19*2R <400> 10 tatcactttc cataaaagca ag 22 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CYP2C19*3F <400> 11 ccaatcattt agcttcaccc 20 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CYP2C19*3R <400> 12 acttcagggc ttggtcaata 20 <210> 13 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> CYP2C19*17F <400> 13 gcccttagca ccaaattctc t 21 <210> 14 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> CYP2C19*17R <400> 14 cacctttacc atttaacccc c 21 <210> 15 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> CYP4F2*3F <400> 15 attgcgggga gaaaaggtc 19 <210> 16 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> CYP4F2*3R <400> 16 ctttgaggga ggtgatgttg g 21 <210> 17 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CYP2C9*3F <400> 17 gacttaccca tgcccctttg 20 <210> 18 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CYP2C9*3R <400> 18 atctggagaa cacacactgc 20 <210> 19 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> CYP2C9*13F <400> 19 gtagagacgt ggtatcacct tg 22 <210> 20 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> CYP2C9*13R <400> 20 cagccagtgg gaaaatgc 18 <210> 21 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> CYP2C9*14F <400> 21 ttacagagct cctcgggcag a 21 <210> 22 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> CYP2C9*14R <400> 22 ggtaatggga aaaacactgc t 21 <210> 23 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> VKORC1-1639F <400> 23 acagcatctg gagagggagg ag 22 <210> 24 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> VKORC1-1639R <400> 24 gcattgccct gacacctagt gg 22 <210> 25 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> HLA-B_AF <400> 25 ccggggtccc agttctaaag 20 <210> 26 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> HLA-B_AR <400> 26 tccgatgacc acaactgcta 20 <210> 27 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> G6PD_F2-2 <400> 27 gaacacaagg cacgggaggt 20 <210> 28 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> G6PD_R2-2 <400> 28 ggaccagtgc gtgagtgtct ca 22 <210> 29 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> HLA-DRB1_F <400> 29 cctgtggcag ggtaagtata 20 <210> 30 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> HLA-DRB1_R <400> 30 cccgtagttg tgtctgcaca c 21 <210> 31 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> HLA-DQA1_F <400> 31 gcagggtttg gtttgggtg 19 <210> 32 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> HLA-DQA1_R <400> 32 tctggtttcc tgaaggagra aca 23 <210> 33 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> HGF_F <400> 33 cagtgccttc ccaaccattc cctta 25 <210> 34 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> HGF_R <400> 34 atccactcac ggatttctgt tgtgtttc 28 <210> 35 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TPMT *6 E8F <400> 35 cttccttccc tgccttttgt 20 <210> 36 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TPMT *6 E8R <400> 36 cccaacaact ttacctggat g 21 <210> 37 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TPMT *3C E10F <400> 37 cacccagcca attttgagta 20 <210> 38 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> TPMT *3C E10R <400> 38 cctcaaaaac atgtcagtgt ga 22 <210> 39 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> NUDT15_F <400> 39 ttgtatagcc aagcaaatgc aaagc 25 <210> 40 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> NUDT15_R <400> 40 tctgtgtctg gaatacaatt caatgac 27 <210> 41 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> DPYD_rs3918290_F <400> 41 actctttcat caggacattg tgacaa 26 <210> 42 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> DPYD_rs3918290_R <400> 42 atttcatttc tcttccttct gcttatg 27 <210> 43 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> DPYD_rs67376798_F <400> 43 agtctcccaa gttaatataa tgcaacatt 29 <210> 44 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> DPYD_rs67376798_R <400> 44 tataagggtg acaggacaga aagatgt 27 <210> 45 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> UGT1A1*28Frag_F <400> 45 cttggtgtat cgattggttt ttg 23 <210> 46 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> UGT1A1*28Frag_R(27mer) <400> 46 ggcgcctttg ctcctgccag aggttcg 27 <210> 47 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> SLCO1B1_g.388_F_26mer <400> 47 ggtgcaaata aaggggaata tttctc 26 <210> 48 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> SLCO1B1_g.388_R_30mer <400> 48 gtatacattt aattacatct ctaaaaattg 30 <210> 49 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> OATPC 521 FBP <400> 49 cccagtctca ggtatgtatt 20 <210> 50 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> OATPC 521 RP <400> 50 tgtaagaaag ccccaatggt a 21 <210> 51 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> BCRP Q141K E5F <400> 51 ggttcatcat tagctagaac tttacc 26 <210> 52 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> BCRP Q141K E5R <400> 52 tggaaagcaa ccatttttga 20 <210> 53 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> SLC14A2_rs1123617F <400> 53 gggataaatg cctcacgctc ata 23 <210> 54 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> SLC14A2_rs1123617R <400> 54 aggaatcagc aggtggagtg ttg 23 <210> 55 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> SLC14A2_rs3745009F <400> 55 gtcatcaatg aagtatcaat gccctc 26 <210> 56 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> SLC14A2_rs3745009R <400> 56 acgcttcttt gtgtttgggg ag 22 <210> 57 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> C11orf65_F <400> 57 gagaaggcag taaagtgaag gaatacag 28 <210> 58 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> C11orf65_R <400> 58 cgccccagcc tctgaagtag 20 <210> 59 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> ACE_1st_PCR_F <400> 59 gactgctgag gccctgcagg 20 <210> 60 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> ACE_1st_PCR_R <400> 60 ggacgtggcc atcacattcg 20 <210> 61 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> COQ2_F <400> 61 gcccagcatt tagcatagca ctt 23 <210> 62 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> COQ2_R <400> 62 ctgctgcaat tcctagcacg aa 22 <210> 63 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> KCNJ11_rs5219_F-2 <400> 63 cgagaggact ctgcagtgag 20 <210> 64 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> KCNJ11_rs5219_R-2 <400> 64 gcttgctgaa gatgagggtc 20 <210> 65 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> NEDD4L_rs4149601_F <400> 65 acgacttccg catactcttc aga 23 <210> 66 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> NEDD4L_rs4149601_R <400> 66 ccacttagga aaaccaatca ggc 23 <210> 67 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> ADRA1A_F <400> 67 aattggcttg ctggctttca a 21 <210> 68 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> ADRA1A_R <400> 68 gccatccgtc ttggagatcc 20 <210> 69 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> AGTR1_F <400> 69 gccaaatccc actcaaacct ttc 23 <210> 70 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> AGTR1_R <400> 70 caggacaaaa gcaggctagg gag 23 <210> 71 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> GNB3_rs5443_F <400> 71 cctcagttct tccccaatgg a 21 <210> 72 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> GNB3_rs5443_R <400> 72 ccttacccac acgctcagac ttc 23 <210> 73 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> MMP3_rs3025058_F <400> 73 caggctggtt tcgagctctg g 21 <210> 74 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> MMP3_rs3025058_R <400> 74 tgggctccac tgtttcttcc tg 22 <210> 75 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PTGS2_rs20417_F <400> 75 atcgccgctt cctttgtcca 20 <210> 76 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> PTGS2_rs20417_R <400> 76 tcgggacaga ctggggcg 18 <210> 77 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> 1758 seqF(T0) <400> 77 cgccttcgcc aaccactcc 19 <210> 78 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> 100 seqF (T0) <400> 78 caacgctggg ctgcacgcta c 21 <210> 79 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> 1611 seqR (T10) <400> 79 tttttttttt gggcccatag cgcgccagga 30 <210> 80 <211> 38 <212> DNA <213> Artificial Sequence <220> <223> 2573 seqF (T16) <400> 80 tttttttttt tttttttggg acccagccca gccccccc 38 <210> 81 <211> 39 <212> DNA <213> Artificial Sequence <220> <223> 2988 seqF (T20) <400> 81 tttttttttt tttttttttt agtgcagggg ccgagggag 39 <210> 82 <211> 45 <212> DNA <213> Artificial Sequence <220> <223> 3877 seqR (T25) <400> 82 tttttttttt tttttttttt tttttctggg catccaggaa gtgtt 45 <210> 83 <211> 50 <212> DNA <213> Artificial Sequence <220> <223> 4125 seqF (T28) <400> 83 tttttttttt tttttttttt tttttttttt cagcttctcg gtgcccactg 50 <210> 84 <211> 55 <212> DNA <213> Artificial Sequence <220> <223> 2850 seqR (T35) <400> 84 tttttttttt tttttttttt tttttttttt tttttcaggt cagccaccac tatgc 55 <210> 85 <211> 60 <212> DNA <213> Artificial Sequence <220> <223> 1887 seqR (T40) <400> 85 tttttttttt tttttttttt tttttttttt tttttttttt agggaggcga tcacgttgct 60 60 <210> 86 <211> 65 <212> DNA <213> Artificial Sequence <220> <223> DelDup_seqR (T45) <400> 86 tttttttttt tttttttttt tttttttttt tttttttttt tttttctcgt cactggtcag 60 gggtc 65 <210> 87 <211> 70 <212> DNA <213> Artificial Sequence <220> <223> 1846 seqF (T50) <400> 87 tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt acccgcatct 60 cccaccccca 70 <210> 88 <211> 41 <212> DNA <213> Artificial Sequence <220> <223> CYP2C19*2SeqF(T20) <400> 88 tttttttttt tttttttttt cactatcatt gattatttcc c 41 <210> 89 <211> 52 <212> DNA <213> Artificial Sequence <220> <223> CYP19*3seqR(T28) <400> 89 tttttttttt tttttttttt ttttttttca aaaaacttgg ccttacctgg at 52 <210> 90 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> CYP2C19*17seqF(T0) <400> 90 tttcaaattt gtgtcttctg ttctcaaag 29 <210> 91 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> CYP4F2*3 seqF(T4) <400> 91 ttttggaacc catcacaacc cagct 25 <210> 92 <211> 57 <212> DNA <213> Artificial Sequence <220> <223> CYP2C9*2seqR(T37) <400> 92 tttttttttt tttttttttt tttttttttt tttttttgcg ggcttcctct tgaacac 57 <210> 93 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> CYP2C9*3seqR(T6) <400> 93 ttttttgggc aggctggtgg ggagaaggtc aa 32 <210> 94 <211> 43 <212> DNA <213> Artificial Sequence <220> <223> CYP2C9*13seqF(T21) <400> 94 tttttttttt tttttttttt tgtgaaggaa gccctgattg atc 43 <210> 95 <211> 49 <212> DNA <213> Artificial Sequence <220> <223> CYP2C9*14seqF(T19)_30mer <400> 95 tttttttttt tttttttttg caatggaaag aaatggaagg agatccggc 49 <210> 96 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> VKORC1 -1639 seqR(T2) <400> 96 tttaggcgtg agccaccgca cc 22 <210> 97 <211> 63 <212> DNA <213> Artificial Sequence <220> <223> HLA-B_*57_667seqF(T33) <400> 97 tttttttttt tttttttttt tttttttttt tttagttgag gccaaaatcc ccgcgggttg 60 gtc 63 <210> 98 <211> 60 <212> DNA <213> Artificial Sequence <220> <223> HLA-B_*58_384seqF(T36) <400> 98 tttttttttt tttttttttt tttttttttt ttttttcagg aggggccgga gtattgggac 60 60 <210> 99 <211> 65 <212> DNA <213> Artificial Sequence <220> <223> DelDup_seqR (T45) <400> 99 tttttttttt tttttttttt tttttttttt tttttttttt tttttctcgt cactggtcag 60 gggtc 65 <210> 100 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> HLA-DRB1_8240F <400> 100 gagtaccggg cggtgacgga gct 23 <210> 101 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> HLA-DQA1_91F <400> 101 ggcattgtgg gtgagtgc 18 <210> 102 <211> 31 <212> DNA <213> Artificial Sequence <220> <223> HGF_473F <400> 102 ttttttttgc tgggaaataa gaggaggaga c 31 <210> 103 <211> 34 <212> DNA <213> Artificial Sequence <220> <223> TPMT*6 seqF(T14) <400> 103 tttttttttt ttttcctggg aaagaagttt cagt 34 <210> 104 <211> 40 <212> DNA <213> Artificial Sequence <220> <223> TPMT*3C seqF(T15)_25mer <400> 104 tttttttttt tttttattga ctgtcttttt gaaaagttat 40 <210> 105 <211> 43 <212> DNA <213> Artificial Sequence <220> <223> G6PD_03(rs5030872)_seqF(T20)_25mer <400> 105 tttttttttt tttttttttt cgggagggac ctgcagagct ctg 43 <210> 106 <211> 48 <212> DNA <213> Artificial Sequence <220> <223> G6PD_04(rs5030868)_seqF(T28) <400> 106 tttttttttt tttttttttt ttttttttcc ggctgtccaa ccacatct 48 <210> 107 <211> 55 <212> DNA <213> Artificial Sequence <220> <223> NUDT15_seqR(T31)_25mer <400> 107 tttttttttt tttttttttt tttttttttt tcatagcctt gttcttttaa acaac 55 <210> 108 <211> 58 <212> DNA <213> Artificial Sequence <220> <223> DPYD_rs3918290_seqF(T38) <400> 108 tttttttttt tttttttttt tttttttttt ttttttttag gctgactttc cagacaac 58 <210> 109 <211> 59 <212> DNA <213> Artificial Sequence <220> <223> DPYD_rs67376798_seqF(T34)_25mer <400> 109 tttttttttt tttttttttt tttttttttt ttttgtagag caagttgtgg ctatgattg 59 <210> 110 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> C11orf65 seqF(T0) <400> 110 attttttatc cgctctgaca 20 <210> 111 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> OATPC_388_SeqR(T2) <400> 111 ttgtcgatgt tgaattttct gatgaat 27 <210> 112 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> OATPC_c.521_SeqR(T7)_30mer <400> 112 caatttttag agatgtaatt aaatgtatac 30 <210> 113 <211> 44 <212> DNA <213> Artificial Sequence <220> <223> BCRP141K SeqF(T19) <400> 113 tttttttttt tttttttttc actctgacgg tgagagaaaa ctta 44 <210> 114 <211> 48 <212> DNA <213> Artificial Sequence <220> <223> ACE seqF(T28) <400> 114 tttttttttt tttttttttt ttttttttcc acataaaagt gactgtat 48 <210> 115 <211> 55 <212> DNA <213> Artificial Sequence <220> <223> COQ2 seqF(T35) <400> 115 tttttttttt tttttttttt tttttttttt tttttctcaa gaaattgatg tgagc 55 <210> 116 <211> 60 <212> DNA <213> Artificial Sequence <220> <223> KCNJ11 seqF(T30)_30mer <400> 116 tttttttttt tttttttttt tttttttttt gtgctgacac gcctggcaga ggaccctgcc 60 60 <210> 117 <211> 63 <212> DNA <213> Artificial Sequence <220> <223> NEDD4L seqF(T43) <400> 117 tttttttttt tttttttttt tttttttttt tttttttttt tttagacgtc tcgcatttga 60 gca 63 <210> 118 <211> 69 <212> DNA <213> Artificial Sequence <220> <223> SLC14A2_rs1123617_seqF(T47) <400> 118 tttttttttt tttttttttt tttttttttt tttttttttt tttttttaca gcttttgaga 60 gccatcccc 69 <210> 119 <211> 74 <212> DNA <213> Artificial Sequence <220> <223> SLC14A2_rs3745009_seqF(T52) <400> 119 tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt ttgctcctga 60 cgaccaataa cccc 74 <210> 120 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> ADRA1A seqR(T7) <400> 120 tttttttttt ggaagactgc tttctgc 27 <210> 121 <211> 48 <212> DNA <213> Artificial Sequence <220> <223> AGTR1 seqF(T28) <400> 121 tttttttttt tttttttttt ttttttttca cttcactacc aaatgagc 48 <210> 122 <211> 41 <212> DNA <213> Artificial Sequence <220> <223> GNB3 seqR(T21) <400> 122 tttttttttt tttttttttt tccactgagg gagaaggcca c 41 <210> 123 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> MMP3 seqR(T0) <400> 123 ggttctccat tcctttgatg gggggaaaaa 30 <210> 124 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PTGS2 seqF(T0) <400> 124 gaggagaatt tacctttccc 20

Claims (10)

In the multi-speed drug gene diagnosis kit for predicting drug side effects and personalized drug treatment for cancer and chronic disease-related multi-prescription drugs,
Including the primers of SEQ ID NO: 1 to 124,
ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1,*2,*3,*17, CYP2C9 *1,*2,*3,*13,*14, CYP2D6 ,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN (duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs5219, MMP3 rs30L15 rs5219, MMP3 rs30L rs5219, MMP3 rs30L15 rs5219, MMP3 rs30L , PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A, *1B, *5, *15, TPMT *1, *3C, *6, UGT1A1 *1, *28 and at least one selected from the group consisting of VKORC1 rs9923231 A kit for diagnosing multiple high-speed drug genes for predicting drug side effects and personalized drug treatment for cancer and chronic disease-related multi-prescription drugs, characterized in determining the presence or absence of mutations. The method of claim 1,
The kit includes 2X PCR Amplification Mix, 2X Multiplex PCR Amplification Mix, amplification primer mix 1 (APM1) consisting of primers of SEQ ID NO: 1 to SEQ ID NO: 4, amplification primer mix 2 consisting of primers of SEQ ID NO: 5 to SEQ ID NO: 6 (APM2), amplification primer mix 3 consisting of primers of SEQ ID NOs: 7 to 8 (APM3), amplification primer mix 4 (APM4) consisting of primers of SEQ ID NOs: 9 to 24, primers of SEQ ID NOs: 25 to 28 Consisting amplification primer mix 5 (APM5), the amplification primer mix 6 (APM6) consisting of the primers of SEQ ID NOs: 29 to 44, the amplification primer mix 7 (APM7) consisting of the primers of SEQ ID NOs: 45 to 46, SEQ ID NO: 47 to Amplification primer mix 8 (APM8) consisting of primers of 56, amplification primer mix 9 (APM9) consisting of primers of SEQ ID NOs: 57 to 66, amplification primer mix 10 (APM10) consisting of primers of SEQ ID NOs: 67 to 76, Snapshot multiplex reagent (SNaPshot MR), snapshot primer mix 1 (SPM1) consisting of primers of SEQ ID NOs: 77-87, snapshot primer mix 2 (SPM2) consisting of primers of SEQ ID NO: 88-99, SEQ ID NO: Snapshot primer mix 3 (SPM3) consisting of primers of 100 to 109, Snapshot primer mix 4 (SPM4) consisting of primers of SEQ ID NO: 110 to 119, Snapshot primer mix consisting of primers of SEQ ID NO: 120 to 124 5 (SPM5), a kit characterized in that it comprises a control DNA and nuclease-free water. The method of claim 1,
The kit is characterized in that it further comprises Shrimp Alkaline Phosphatase (SAP) and/or Exonuclease 1 (Exo)). The method of claim 1,
The kit, characterized in that the side effects are selected from the group consisting of Stevens-Johnson syndrome, toxic epidermal necrosis dissolution, irritability syndrome, severe liver injury and muscle disease. The method of claim 1,
The drugs are voriconazole, piroxicam, aspirin, celecoxib, diclofenac, tramadol, ibuprofen, omeprazole, esomeprazole, lansoprazole, pantoprazole, rabeprazole, simbassatin, serivastatin, pravastatin, rosuba Statin, atorvastatin, clopidogrel, flecainide, propaphenone, nifedipine, captopril, quinacryl, enalapril, lisinopril, telmisartan, losartan, metoprolol, irbesartan, candesartan, carve Dirol, hydrochlorothiazide, warfarin, glimepiride, glibenclamide, gliclazide, metformin, allopurinol, lapatinib, tamoxifen, cisplatin, mercaptopurine, pazopanib, capecitabine, fluorouracil, irinotecan, Kit, characterized in that at least one selected from the group consisting of azathioprine, rasburicase and similar drugs. A method for diagnosing multiple high-speed drug genes for predicting drug side effects and personalized drug treatment for cancer and chronic disease-related multi-prescription drugs, comprising the following steps:
(a) Using the kit according to claim 1, ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1, *2, *3, *17, CYP2C9 *1, *2, *3,*13,*14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,* XN (Duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T , KCNJ11 rs5219, MMP3 rs325058, NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*1B,*5,*15, TPMT *1,*3C,*6*28, UGT And determining a SNP capable of specifically detecting at least one genotype selected from the group consisting of VKORC1 rs9923231.
(b) ABCG2, ACE, ADRA1A, AGTR1, C11orf65, COQ2, CYP2C19, CYP2C9, CYP2D6, CYP4F2, DPYD, G6PDGNB3, HLA-B, HLA-DQA1, HLA-DRB1, Internal Control HGF, NEDD15, NUDT15, PTGS2, NUDT15 , SLC14A2, SLCO1B1, TPMT, UGT1A1, and performing a multiple polymerase chain reaction to amplify one or more genes selected from the group consisting of VKORC1; And
(c) performing a snapshot (SNaPshot) analysis using multiple single base primers to analyze the SNP for the product to confirm the genotype. The method of claim 6,
In the step (b), the multiple polymerase chain reaction is characterized in that carried out using one or more primers selected from the group consisting of primers of SEQ ID NOs: 1 to 76. The method of claim 7,
The multiple polymerase chain reaction specifically amplifies CYP2D6 using SEQ ID NOs: 1 to 8, specifically amplifies CYP2C19 using SEQ ID NOs: 9 to 14, and CYP4F2 using SEQ ID NOs: 15 to 16. Specific amplification, specific amplification of the CYP2C9 gene using SEQ ID NOs: 17 to 22, specific amplification of the VKORC1 gene using SEQ ID NOs: 23 to 24, and HLA- B gene was specifically amplified, G6PD gene was specifically amplified using SEQ ID NOs: 27 to 28, HLA-DRB1 gene was specifically amplified using SEQ ID NOs: 29 to 30, and SEQ ID NOs: 31 to 32 Amplify the HLA-DQA1 gene specifically using, and specifically amplify the amplification quality control gene HGF using SEQ ID NOs: 33 to 34, and specifically amplify the TPMT gene using SEQ ID NOs: 35 to 38. , Specific amplification of the NUDT15 gene using SEQ ID NOs: 39 to 40, specific amplification of the DPYD gene using SEQ ID NOs: 41 to 44, and specific amplification of the UGT1A1 gene using SEQ ID NOs: 45 to 46 And, the SLCO1B1 gene was specifically amplified using SEQ ID NOs: 47 to 50, and the ABCG2 gene was specifically amplified using SEQ ID NOs: 51 to 52, and the SLC14A2 gene was specifically amplified using SEQ ID NOs: 53 to 56. Amplification, specific amplification of the C11orf65 gene using SEQ ID NOs: 57 to 58, specific amplification of the ACE gene using SEQ ID NOs: 59 to 60, and specific COQ2 gene using SEQ ID NOs: 61 to 62 And amplified with SEQ ID NOs: 63 to 64, specifically amplifying the KCNJ11 gene, and specifically amplifying the NEDD4L gene using SEQ ID NOs: 65 to 66, and ADRA1A using SEQ ID NOs: 67 to 68 Amplify the gene specifically, specifically amplify the AGTR1 gene using SEQ ID NOs: 69 to 70, specifically amplify the GNB3 gene using SEQ ID NOs: 71 to 72, and use SEQ ID NOs: 73 to 74 A method, characterized in that the MMP3 gene is specifically amplified and the PTGS gene is specifically amplified using SEQ ID NOs: 75 to 76. The method of claim 6,
In the step (c), the snapshot analysis is characterized in that performed using one or more primers selected from the group consisting of the primers of SEQ ID NOs: 77 to 124. The method of claim 6,
In the step (c), genotyping is, ABCG2 rs2231142, ACE rs1799752, ADRA1A rs1048101, AGTR1 rs5186, C11orf65 rs11212617, COQ2 rs4693075, CYP2C19 *1, *2, *3, *17, CYP2C9 *1, *2, * 3,*13,*14, CYP2D6 *1,*2,*4,*5 (defect),*10,*14,*18,*21,*41,*49,*52,*60,*XN (Duplicate), CYP4F2 *1,*3, DPYD rs3918290, rs67376798, G6PD rs5030872, rs5030868, GNB3 rs5443, HLA-B *5701,*5801, HLA-DQA1*02, HLA-DRB1*07, Internal Control HGF 473T, KCNJ11 rs5219, MMP3 rs3025058, NEDD4L rs4149601, NUDT15 rs116855232, PTGS2 rs20417, SLC14A2 rs1123617, rs3745009, SLCO1B1 *1A,*1B,*5,*15, TPMT *1,*1A1 *1,*28 and UGT VKORC1 rs9923231, characterized in that to identify one or more mutations selected from the group consisting of.

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