HK1140234A - Methods for selecting medications - Google Patents

Description

Method for selecting therapeutic agents

Technical Field

The present invention relates to a method of selecting a therapeutic drug (e.g., psychotherapeutic drug) for a patient, and more particularly to selecting a therapeutic drug for a patient based on the genotype of a gene encoding a drug-metabolizing enzyme and a gene encoding a product involved in, for example, neurotransmission.

SUMMARY

The present invention is based on the identification of a group of genes with polymorphisms relevant for the diagnosis of psychosis and pharmacological response to therapeutic drugs. Thus, the methods of the invention allow for the determination of a patient's genotype and the selection of an appropriate therapeutic agent for the patient based on that genotype. The method of the present invention allows for the integration of the output information from multiple genotypic assessments, providing important and improved clinical information based on which therapeutic agents are selected and administered. Thus, the methods of the present invention provide a rational method for identifying therapeutic agents that will achieve the optimal response in a patient.

In one aspect, the invention features a method of selecting a psychotherapeutic drug (e.g., an antidepressant, antipsychotic, anxiolytic-sedative, or mood stabilizer) for a patient. The method comprises providing a genotype for a set of genes in said patient, wherein said set of genes comprises at least 3 cytochrome P450 genes and a 5-hydroxytryptamine transporter gene and selecting a psychotherapeutic drug based on said genotype. The at least 3 cytochrome P450 genes may encode CYP2D6, 1A2 and 2C19 and providing the genotype of the patient comprises determining whether the patient comprises a CYP1A 2A 1A or 1A2 A3 allele, CYP2C19 a 1A, 2C19 a 1B, or 2C19 a 2A allele and CYP2D 6a, 2D 6a 2, 2D 6N, 2D 6A3, 2D 64, 2D 65, 2D 6a 6, 2D 67, 2D 68, 2D6 10, 2D6 12 or 2D 6a 17 allele. The method may further comprise determining whether the patient comprises the 2D6 x 41 allele.

The set of genes may also include a 5-hydroxytryptamine receptor 2A gene. For example, the set of genes may include cytochrome P450 genes encoding CYP2D6, 2C19, and 1A2, the 5-hydroxytryptamine transporter gene, and the 5-hydroxytryptamine receptor 2A gene. Selecting the psychotherapeutic drug may comprise correlating the genotype of the cytochrome P450 gene with the ability of each cytochrome P450 enzyme encoded by each cytochrome P450 gene to metabolize the psychotherapeutic drug; and correlating the genotype of the 5-hydroxytryptamine transporter gene and the 5-hydroxytryptamine receptor gene with the patient's ability to respond to the psychotherapeutic drug.

The set of genes may include 4 cytochrome P450 genes (e.g., genes encoding CYP2D6, 2C19, 3a4, and 1a 2). Providing the genotype of the patient may comprise determining whether the patient comprises a CYP2D6 x 2, 2D6 x 3, 2D6 x 4, 2D6 x 10 or 2D6 x 17 allele, CYP2C19 x 2A, 2C19 x 2B, 2C19 x 3, 2C19 x 4, 2C19 x 5A, 2C19 x 5B, 2C19 x 6, 2C19 x 7 or 2C 465 x 8 allele, CYP3a4 x 1B, 3a4 x 2, 3a4 x 5, 3a4 x 6, 3a4 x 12, 3a4 x 13, 3a 695A 2x 15A, 3a4 x 5, 3a4 x 6, 3a 863 a and a 8427 x 861 allele.

The set of genes may include at least 5 cytochrome P450 genes (e.g., CYP1a1, 1a2, 2D6, 2C19, and 3a 4). Providing the patient's genotype may comprise determining whether the patient comprises a CYP1A 1x 1A, 1A 1x 2, 1A 1x 3 or 1A 1x 4 allele, a CYP1A 2x 1A or 1A 2x 3 allele, a CYP2C19 x 1A, 2C19 x 1B or 2C19 x 2A allele, a CYP2D6 x 1A, 2D6 x 2, 2D6 x 2N, 2D6 x 3, 2D6 x 4, 2D6 x 5, 2D6 x 6, 2D6 x 7, 2D6 x 8, 2D6 x 10, 2D6 x 12, 2D6 x 6, 2D6 x 7, 2D6 x 8, 2D6 x 10, 2D 863 x 863, or a 82863 a allele.

The set of genes may also include a plurality of dopamine receptor genes (e.g., dopamine receptor genes encoding dopamine receptors D1, D2, D3, D4, D5, and D6), a plurality of 5-hydroxytryptamine receptor genes (e.g., 5-hydroxytryptamine receptor genes encoding 5-hydroxytryptamine receptors 1A, 1B, 1D, 2A, or 2C), tryptophan hydroxylase genes, and/or catechol-o-methyltransferase genes. The 5-hydroxytryptamine receptor gene 2A is particularly useful. In some embodiments, the set of genes includes CYP2D6, 2C19, 3a4, and 1a2 genes, 5-hydroxytryptamine receptor genes, and 5-hydroxytryptamine receptor 2A genes.

In some embodiments, the set of genes includes at least 10 cytochrome P450 genes (e.g., genes encoding CYP1a1, 1a2, 1B1, 2A6, 2B6, 2C8, 2C9, 2C18, 2C19, 2D6, 2E1, 3a4, and 3a 5). Determining the genotype of the patient may comprise determining whether the patient comprises a CYP1A 1x 1A, 1A 1x 2, 1A 1x 3, or 1A 1x 4 allele; 1A 2x 1A or 1A 2x 3 allele; CYP1B 1x 1, 1B 1x 2, 1B 1x 3, 1B 1x 4, 1B 1x 11, 1B 1x 14, 1B 1x 18, 1B 1x 19, 1B 1x 20, or 1B 1x 25 alleles; CYP2a6 x 1A, 2a6 x 1B, 2a6 x 2 or 2a6 x 5 allele; CYP2B6 x 1, 2B6 x 2, 2B6 x 3, 2B6 x 4, 2B6 x 5, 2B6 x 6, or 2B6 x 7 alleles; CYP2C8 x 1A, 2C8 x 1B, 2C8 x 1C, 2C8 x 2, 2C8 x 3, or 2C8 x 4 alleles; CYP2C9 x 1, 2C9 x 2, 2C9 x 3 or 2C9 x 5 allele; CYP2C18 m1 or 2C18 m2 alleles; CYP2C19 x 1A, 2C19 x 1B or 2C19 x 2A alleles; CYP2D 6a, 2D 6a 2, 2D 6a 2N, 2D 6a3, 2D 6a 4, 2D 6a 5, 2D 6a 6, 2D 6a 7, 2D 6a 8, 2D 6a 10, 2D 6a 12, 2D 6a 17 or 2D 6a 35 allele; CYP2E 1x 1A, 2E 1x 1C, 2E 1x 1D, 2E 1x 2, 2E 1x 4, 2E 1x 5 or 2E 1x 7 alleles; CYP3a4 x 1A or 3a4 x 1B allele; and CYP3a5 x 1A, 3a5 x 3, 3a5 x 5 or 3a5 x 6 alleles.

In another aspect, the invention features an article that includes a substrate. The substrate comprises a plurality of discrete regions, wherein each region comprises a different population of nucleic acid molecules, wherein the different populations of nucleic acid molecules independently comprise nucleic acid molecules for detecting: CYP1A 2x 1A and 1A 2x 3 alleles; CYP2C19 x 1A, 2C19 x 1B, and 2C19 x 2A alleles; CYP2D 6a, 2D 62, 2D 6N, 2D 63, 2D 64, 2D 65, 2D 66, 2D 67, 2D 68, 2D6 10, 2D6 12, 2D6 17 and 2D 635 alleles; and 5HTT promoter repeat and exon 2 variable repeat alleles. The population of nucleic acid molecules may further include nucleic acid molecules for detecting one or more of the following alleles: CYP1A 1x 1A, 1A 1x 2, 1A 1x 3 and 1A 1x 4 alleles; CYP1B1 × 1, 1B1 × 2, 1B1 × 3, 1B1 × 4, 1B1 × 11, 1B1 × 14, 1B1 × 18, 1B1 × 19, 1B1 × 20 and 1B1 × 25 alleles, CYP2A6 × 1A, 2A6 × 1B, 2A6 × 2 and 2A6 × 5 alleles, CYP2B6 × 1, 2B6 × 2, 2B6 × 3, 2B6 × 4, 2B6 × 5, 2B6 × 6 and 2B6 × 7 alleles, CYP2C8 × 1A, 2C8 × 1B, 2C8 × 1C, 2C8 × 2, 2C8 × 3 and 2C8 × 4 alleles, CYP2C9 × 1, 2C9 × 2, 2C9 × 3 and 2C9 × 5 alleles, CYP2C18 × m1 and 2C18 × 2 alleles, CYP2C19 × 1A, 2C19 × 1B and 2C19 × 2A alleles, CYP2E1 × 1A, 2E1 × 1C, 2E 7 × 1D, 2E1 × 2, 2E1 × 1A and 1A1 alleles; DAT140bp VNTR and 10 repeat allele; and CYP3a5 x 1A, 3a5 x 3, 3a5 x 5 and 3a5 x 6 alleles. The population may also include nucleic acid molecules for detecting TPH a218C, a779C, G5806T, a6526G, and (CT) m (ca) n (CT) p alleles.

The invention also features a method of constructing a database for selecting a therapeutic agent for a patient. The method includes receiving, in a computer system, a plurality of genotypes of a set of genes, the set of genes including a CYP2D6 gene, a CYP2C19 gene, a CYP1a2 gene, a 5-hydroxytryptamine transporter gene, and a 5-hydroxytryptamine receptor 2A gene; receiving a plurality of therapeutic profiles (mediationprofiles) assigned based on the genotype; and storing a plurality of genotypes and therapeutic drug histories such that each therapeutic drug history is associated with one of the genotypes. The at least one treatment agent may identify the treatment agent and the treatment agent may be placed in one of a plurality of categories included in the treatment agent. Such categories may be selected from the group consisting of: safe therapeutic agents to use, therapeutic agents that should be used with caution, therapeutic agents that should be closely monitored while in use, therapeutic agents that should be avoided, and combinations thereof. Therapeutic agents the patient's genotype may be identified with a range of possible therapeutic agents.

In another aspect, the invention features a computer program product including executable instructions that when executed cause a processor to operate. The operations may include: receiving a plurality of genotypes of a set of genes, wherein the set of genes comprises a CYP2D6 gene, a CYP2C19 gene, a CYP1A2 gene, a 5-hydroxytryptamine transporter gene, and a 5-hydroxytryptamine receptor 2A gene; receiving a plurality of therapeutic drug regimens specified based on the genotype; and storing the genotypes and the therapeutic drug histories to associate each therapeutic drug history with one of the genotypes.

The invention also features a method of selecting a therapeutic agent for a patient. The method includes receiving in a computer system a genotype for a set of genes in a patient, wherein the set of genes includes a CYP2D6 gene, a CYP2C19 gene, a CYP1a2 gene, a 5-hydroxytryptamine transporter gene, and a 5-hydroxytryptamine receptor 2A gene; identifying a course of therapeutic drugs associated with the genotype of the patient in a database comprising a plurality of courses of therapeutic drugs associated with genotypes; and outputting the identified therapeutic drug history in response to receiving the patient's genotype. The user may enter the patient's genotype in a computer system or the patient's genotype may be received directly from the device for determining the patient's genotype.

The therapeutic agent profile may include a ranking of a plurality of therapeutic agents, e.g., based on a particular cofactor. The method may include adjusting the ranking prior to outputting the identified course of treatment (e.g., based on receiving a genotypic polymorphism carried by the patient or based on receiving a clinical response for the patient). The clinical response is by the patient's family members.

In another aspect, the invention features a computer program product including executable instructions that, when executed, cause a processor to perform operations including: receiving genotypes of a set of genes of the patient, wherein the set of genes comprises a CYP2D6 gene, a CYP2C19 gene, a CYP1A2 gene, a 5-hydroxytryptamine transporter gene, and a 5-hydroxytryptamine receptor 2A gene; identifying a course of therapeutic drugs associated with the genotype of the patient in a database comprising a plurality of courses of therapeutic drugs associated with genotypes; and outputting the identified therapeutic drug history in response to receiving the patient's genotype.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims.

Drawings

FIG. 1 is a block diagram of a computer system 100 according to one embodiment.

Fig. 2 is a flow chart of a method 200 of building a database for patient selected therapeutic drugs.

Fig. 3 is a flow chart of a method 300 of selecting a therapeutic agent for a patient.

Detailed description of the invention

In summary, the invention features a method of selecting a therapeutic agent (e.g., a psychotherapeutic agent) for a patient based on the genotype of a gene useful for selecting the therapeutic agent. The genes to be genotyped typically encode products that affect the metabolism of the therapeutic drug or are associated with a better therapeutic response. Algorithms for the initial exclusion of therapeutic drugs based on the problematic (proliferative) genotype of drug metabolizing enzyme genes (e.g., cytochrome P450 genes) can be used. The second phase of the algorithm may begin with a list of possible appropriate therapeutic drugs (e.g., antidepressants) and obtain a further classification of the optimal treatment based on the genotype of the target gene. For example, for antidepressants, a further classification of potential antidepressants may be based on the genotypes of the 5-hydroxytryptamine transporter (5HTTR) and 5-hydroxytryptamine receptor 2A (HTR 2A).

Psychotherapeutic drugs include antipsychotics or neuroleptics, anxiolytics-sedatives, antidepressants or mood-elevating agents (mood-elevating agents) and mood-stabilizing agents (e.g., lithium salt or valproic acid). Non-limiting examples of antipsychotics include tricyclic phenothiazines (e.g., chlorpromazine, trifluoropropazine, thioridazine and mesoridazine, fluphenazine and trifluoperazine), thioxanthenes (e.g., chlorprothixene, clopenthixol, trifluorothixene, pipothioxol and thiothixene), and diphenipine (e.g., loxapine, clozapine, chlorothiapine, methiothepin, zotepine (zotepine), ICI-204,636, fluperlatine and olanzapine); phenylpropenones such as haloperidol, diphenylbutylpiperidines such as fluspirilene, pentafluridol and pimozide, haloperidol decanoate and indolone. Non-limiting examples of anxiolytic-sedatives include benzodiazepines such as chlordiazepoxide, diazepam, oxazepam, clorazepate (clorazepate), lorazepam, pramipepam, alprazolam, and halazepam. Antidepressants or mood stimulants include norepinephrine reuptake inhibitors such as tertiary amine tricyclic antidepressants (e.g., amitriptyline, clomipramine, doxepin, and imipramine) and secondary amine tricyclic antidepressants (e.g., amoxapine, desipramine, maprotiline, protriptyline, and nortriptyline); selective 5-hydroxytryptamine reuptake inhibitors (SSRIs) such as fluoxetine, fluvoxamine, paroxetine, sertraline, citalopram, escitalopram and venlafaxine; atypical antidepressants such as bupropion, nefazodone and trazodone; noradrenergic and specific 5-hydroxytryptamine antidepressants such as mirtazapine; and monoamine oxidase inhibitors such as phenelzine, tranylcypromine, and selegiline.

In some ethnic groups, up to 10% of the adolescent population have a weak association with metabolism of many antidepressant therapeutic drugsThe 2D6 haplotype of (a). See Wong et al (2001)Ann.Acad.Med. Singapore29: 401-406. Clinical genomic testing of these individuals is clearly associated with their treatment and prognosis. In extreme cases, children of the weak metabolic type (metabolizer) and unidentified children have tragic results. These negative case reports included reporting the death of 9 year old boys that were not recognized as weakly metabolic 2D 6. The boy was treated continuously with fluoxetine, although multiple symptoms developed, as these were not recognized to be associated with the extremely high levels of fluoxetine in his serum. Sallee et al (2000)J.Child Adol.Psychiatry10(1): 27-34. Although careful clinical monitoring during administration of low doses of therapeutic drugs to identify undesirable side effects is an alternative clinical strategy for genotyping, genomic detection of multiple genes encoding drug metabolizing enzymes (e.g., cytochrome P450 genes) and other genes of interest (e.g., genes involved in neurotransmission for therapeutic psychotropic drugs) provides a safe method by which potentially dangerous side effects in infected patients can be avoided.

Each group of genes

The method includes obtaining a biological sample from a patient and obtaining the genotype of a set of genes of the patient. Typically the set of genotyped genes includes at least 3 cytochrome P450 genes. The cytochrome P450 gene may be selected from the P450 genes listed in table 1. For example, the at least 3 cytochrome P450 genes can encode CYP2D6, 1a2, and 2C 19. In embodiments where antidepressant drug therapy is selected, the genotypes of the 4 cytochrome P450 genes (e.g., the genes encoding CYP2D6, 2C19, 3a4, and 1a 2) are available. In other embodiments, the genotype of at least 5 cytochrome P450 genes (e.g., genes encoding CYP1a1, 1a2, 2D6, 2C19, and 3a4) can be obtained. In other embodiments, the genotype of at least 10 cytochrome P450 genes (e.g., genes encoding CYP1a1, 1a2, 1B1, 2A6, 2B6, 2C8, 2C9, 2C18, 2C19, 2D6, 2E1, 3a4, and 3a 5) can be obtained. The alleles of each of these cytochrome P450 genes are listed in table 1.

The substrates of CYP2D6 are typically weak bases with a cation binding site located away from the carbon atom to be oxidized. Specifically, substrates for CYP2D6 include amitriptyline, nortriptyline, haloperidol, and desipramine. Venlafaxine is another substrate for CYP2D6 and can be bioconverted primarily to the active metabolite O-desmethylvenlafaxine by the CYP2D6 enzyme. Venlafaxine can also be metabolized to the inactive metabolite N-desmethylvenlafaxine (N-desmethylvenlafaxine), which requires the CYP2D6 enzyme. Some individuals have altered CYP2D6 gene sequences, resulting in a lack of catalytic activity or reduced catalytic activity of the synthesized enzyme. These individuals are poorly metabolized by SSRIs (e.g., venlafaxine) and tricyclic antidepressants (TCAs). Duplication of the functional CYP2D6 gene has also been observed and results in ultra-fast metabolism of SSRIs and other drugs. Individual phenotypes without inactivated polymorphisms, deletions or duplications are extensive drug metabolisms and are labeled CYP2D6 x 1. The CYP2D6 x 2 allele has reduced enzymatic activity due to amino acid substitutions. The CYP2D6 x 3 and x 4 alleles account for nearly 70% of the total defects in the phenotype responsible for the weak metabotropic form. Polymorphisms responsible for CYP2D6 × 3(2549A > del) produce frameshifts in mRNA. Polymorphisms involving the CYP2D6 x 4 allele (1846G > a) perturb mRNA splicing. These changes produce truncated forms lacking catalytic activity for CYP2D 6. Other weak metabolic forms are CYP2D6 x 5, x 10 and x 17. CYP2D6 × 5 was due to complete gene deletion. Polymorphisms CYP2D6 x 10 and x 17 produce amino acid substitutions in CYP2D6 enzymes, reducing their enzymatic activity. All of these polymorphisms are autosomal recessive. Therefore, only individuals who are homozygous or compound heterozygous for these polymorphisms are weakly metabotropic. Heterozygous individuals with one normal gene and one polymorphic gene will have intermediate metabolism between the extensive (normal) and weak metabolic types.

CYP1a2 metabolizes many aromatic and heterocyclic amines, including clozapine and imipramine. The CYP1a 2x 1F allele can lead to high inducibility or increased activity of the product. See Sachse et al (1999)Br.J.Clin.Pharmacol.47: 445-449. CYP2C19 also metabolizes a number of substrates including imipramine, citalopram, and diazepam. CYP2C19 a, 2B, 3, 4, 5A, 5B, 6, 7 and 8 alleles encode products with little or no activity. See Ibenu et al (1999)J. Pharmacol.Exp.Ther.290：635-640。

CYP1a1 may be associated with toxic or allergic reactions due to the production of reactive metabolites outside the liver. CYP3a4 metabolizes a variety of substrates, including alprazolam. CYP1B1 may be associated with toxic or allergic reactions due to extrahepatic production of reactive metabolites and also metabolize steroid hormones (e.g. 17 β -estradiol). Substrates for CYP2a6 and CYP2B6 include valproic acid and bupropion, respectively. Substrates for CYP2C9 include tylosin and antabust (dithiolene). Substrates for CYP2E1 include phenytoin and carbamazepine. A decrease in the activity of one or more cytochrome P450 enzymes may affect one or more other cytochrome P450 enzymes.

TABLE 1

Cytochrome P450 gene

Cytochrome P450 gene	Alleles	Polymorphism
			1A1	*1A*2*3*4	No A2455GT3205CC2453A
1A2	*1A*1F*3	none-164C > AG1042A
			1B1	*1*2*3*4*11*14*18*19	No R48GL432VN453SV57CE281XG365WP379L

Cytochrome P450 gene	Alleles	Polymorphism
				*20*25	E387KR469W
2A6	*1A*1B*2*5	No CYP2a7 translocation to the 3' -end T479A x 1B + G6440T
			2B6	*1*2*3*4*5*6*7	R22CS259CK262RR487CQ172H；K262RQ172H；K262R；R487C
2C8	*1A*1B*1C*2*3*4	none-271C > A-370T > GI269FR 139K; k399RI264M
			2C9	*1*2*3*5	No R144CI359LD360E
2C18	m1m2	T204AA460T
			2C19	*1A*1B*2A*2B*3*4*5(A，B)*6*7*8	No splicing defect of I331V; E92D New stop codon 636G > AGTG initiation codon, 1A > G1297C > T, amino acid Change (R433W)395G > A, amino acid Change (R132Q) IVS5+2T > A, splice Defect 358T > C, amino acid Change (W120R)
2D6	*1A*2*2N*3*4*5	No G1661C, C2850T gene duplication A2549 deletion G1846A gene deletion

Cytochrome P450 gene	Alleles	Polymorphism
				*6*7*8*10*12*17*35	T1707 deletion A2935CG1758TC100TG124AC1023T，C2850TG31A
2E1	*1A*1C，*1D*2*4*5*5*7*7*7	No (6 or 8bp repeats) G1132AG476AG (-1293) CC (-1053) TT (-333) AG (-71) TA (-353) G
			3A4	*1A*1B*2*5*6*12*13*15A*17*18A	No A (-392) G amino acid change (S222P) amino acid change (P218R) frameshift, 831ins A amino acid change (L373F) amino acid change (P416L) amino acid change (R162Q) amino acid change (F189S, decreasing) amino acid change (L293P, increasing)
3A5	*1A*3*5*6	GT12952CG14960A without A698

Usually, for the selection of therapeutic drugs, in addition to the genotype of the gene encoding the drug-metabolizing enzyme, the genotype of other target genes is obtained. The target gene may encode a product that is relevant to the patient's ability to respond to a particular type of therapeutic agent. For example, for selection of antidepressant drug target genes may be the 5-hydroxytryptamine transporter gene and the 5-hydroxytryptamine receptor 2A gene. Thus, in one embodiment, the set of genes can be the CYP2D6 gene, the 1a2 gene, the 2C19 gene, the 5-hydroxytryptamine transporter gene, and the 5-hydroxytryptamine receptor 2A gene. For selection of an anti-stressor (antipsychotic), the target gene may be a dopamine transporter gene.

Table 2 lists alleles of genes of interest that can be genotyped in addition to the gene encoding a drug metabolizing enzyme. For example, a set of genes to be genotyped can include one or more dopamine receptor genes (e.g., genes encoding dopamine receptors D1, D2, D3, D4, D5, and/or D6), dopamine transporter genes, one or more 5-hydroxytryptamine receptor genes (e.g., genes encoding 5-hydroxytryptamine receptors 1A, 1B, 1D, 2A, or 2C), catechol-o-methyltransferase (COMT) genes, or tryptophan hydroxylase genes. In one embodiment, the COMT gene and tryptophan hydroxylase gene are included in the panel along with the cytochrome P450 gene. In other embodiments, one or more dopamine receptor genes, one or more 5-hydroxytryptamine receptor genes, a COMT gene, and a tryptophan hydroxylase gene are evaluated along with a cytochrome P450 gene.

TABLE 2

Gene	Symbol	Polymorphism
			Dopamine transporters	DAT1，SLC6A3	40bpVNTR10 repeat alleles G710A, Q237RC124T, L42F
Dopamine receptor D1	DRD1	DRD1B2T244GC179TG127AT11GC81TT595G，S199AG150T，R50SC110G，T37RA109C，T37P
			Dopamine receptor D2	DRD2	TaqI AA1051G，T35AC932G，S311CC928，P310SG460A，V154I
Dopamine receptor D3	DRD3	BalIMspidRD31Gly/Ser (allele 2) A25G, S9G in exon I

Gene	Symbol	Polymorphism
			Dopamine receptor D4	DRD4	12/13bp insertion/deletion of the 48 repeat 7 repeat allele in exon 3T 581G, V194GC841G, P281A
Dopamine receptor D5	DRD5	T978CL88FA889C，T297PG1252A，V418IG181A，V61MG185C，C62ST263G，R88LG1354A，W455
			Tryptophan hydroxylase	TPH	A218CA779CG-5806TA-6526G 3' UTR middle (CT)m(CA)n(CT)pAllele 194, 115657bp apart from exon
5-hydroxytryptamine transporters	5-HTTR	Promoter repeats (44bp insertion (L)/deletion (S) (L: long form; S: short form), intron 2 variable number (9, 10, 11, or 12); A1815C; G603C; G167C; -3745, T → A (5 ' FR); 3636, T → C (5 ' FR); -3631G → A (5 ' FR); SNP rs25531, A → G (5 ' FR); -1090, A → T (5 ' FR); -1089, A → T (5 ' FR); 482, T → C (5 ' FR); -469, C → T (5 ' FR); -45, C → A (intron 1A); -25, G → A intron 1 UTR-185, A (5 ' FR); A → C (149); C → 303; C → A → 303); intron 2 (6728); exon C), g → A (intron 4); C83T (intron 7); C1149T (exon 8); T204G (intron 8); -131, C → T (intron 11)
			5-hydroxytryptamine receptor 1A	HTR1A	RsaIG815A，G272DG656T，R219LC548T，P551LA82G，I28VG64A，G22SC47T，P16L
5-hydroxytryptamine receptor 1B	HTR1B	G861CG861C，V287VT371G，F124CT655C，F219L

Gene	Symbol	Polymorphism
					A1099G，I367VG1120A，E374K
5-hydroxytryptamine receptor 1D	HTR1D	G506TC173TC794T，S265L
			5-hydroxytryptamine receptor 2A	HTR2A	C74AT102CT516CC1340TC1354T
5-hydroxytryptamine receptor 2C	HTR2C	G796CC10G，L4VG68C，C23S
			Catechol-o-methyltransferase	COMT	G158A (also known as Val/Met) G214TA72SG101CC34SG473A

For example, to select an antidepressant, the patient may be evaluated for the genotype of 3 (e.g., genes encoding 2D6, 2C19, and 1a 2) or 4 cytochrome P450 genes (e.g., genes encoding 2D6, 2C19, 3a4, and 1a 2), the 5-hydroxytryptamine transporter (5HTTR) gene, and the 5-hydroxytryptamine receptor 2A (HTR2A) gene. In particular, it can be determined whether the patient comprises CYP2D6 x 2, # 3, # 4, # 10, # 17 or # 5del alleles, CYP2C19 x 2(A, B), # 3, # 4, # 5(A, B), # 6, # 7, # 8 alleles, CYP1a2 1F alleles, short or long forms of 5-hydroxytryptamine transporter genes and HTR2A T102C polymorphisms. Embodiments that include the 3a4 gene in a panel of genes can determine whether a patient contains the CYP3a4 x 1B, 2, 5, 6, 12, 13, 15A, 17, or 18A allele. Each of these genes affects the metabolism of at least one antidepressant therapeutic drug or is accompanied by a better therapeutic response. As described herein, algorithms have been established based on a set of 6 rules for genotypes of 6 genes (e.g., 2D6 gene, 2C19 gene, 3a4 gene, 1a2 gene, 5HTTR gene, and HTR2A gene). Similarly, an algorithm may be established based on a set of 5 rules for 5 genes (e.g., 2D6 gene, 2C19 gene, 1a2 gene, 5HTTR gene, and HTR2A gene). Based on these algorithms, a patient-based therapeutic drug history is provided for a given patient, enabling clinicians to select acceptable antidepressants without having to trial and error determine whether the patient will respond to or tolerate a particular antidepressant.

Determining genotype

Genomic DNA is commonly used to determine genotype, although mRNA may also be used. Genomic DNA is typically extracted from a biological sample, such as a peripheral blood sample, but can be extracted from other biological samples, including tissues (e.g., oral wall mucosal scrapings or from kidney or liver tissue). Conventional methods can be used to extract genomic DNA from blood or tissue samples, including, for example, phenol extraction. Alternatively, genomic DNA may be extracted in a kit, e.g.Tissue kits (Qiagen, Chatsworth, Calif.),Genomic DNA purification kit (Promega) and a.s.a.p.^TMGenomic DNA isolation kit (Boehringer Mannheim, Indianapolis, IN).

Typically, an amplification step is performed prior to genotyping. For example, Polymerase Chain Reaction (PCR) techniques can be used to free the patient fromThereby obtaining an amplification product. PCR refers to an assay protocol or technique in which a target nucleic acid is enzymatically amplified. Sequence information from or beyond each end of the target region is typically used to design oligonucleotide primers whose sequence is identical to the opposite strands of the template to be amplified. PCR can be used to amplify specific sequences from DNA and RNA, including sequences from total genomic DNA or total cellular RNA. Primers are typically 14 to 40 nucleotides in length, but can vary in length from 10 nucleotides to hundreds of nucleotides. General PCR techniques are described, for example, inPCR Primer：A Laboratory Manual (PCR primer: laboratory Manual)Dieffenbach, C. and Dveksler, G. ed., Coldspring Harbor Laboratory Press, 1995. When RNA is used as a template source, reverse transcriptase can be used to synthesize complementary dna (cdna) strands. Ligase chain reaction, strand displacement amplification, self-sustained sequence amplification or nucleic acid sequence-based amplification may also be used to obtain isolated nucleic acids. See, e.g., Lewis (1992) Genetic Engineering News 12 (9): 1; guatelli et al (1990) Proc.Natl.Acad.Sci.USA 87: 1874-1878; and Weiss (1991) Science 254: 1292-1293.

Primers are typically single-or double-stranded oligonucleotides, 10 to 50 nucleotides in length, capable of being extended when subjected to PCR conditions in combination with mammalian genomic DNA to produce a nucleic acid product corresponding to a target region within a gene. Typically, the PCR product is at least 30 nucleotides in length (e.g., 30, 35, 50, 100, 250, 500, 1000, 1500, or 2000 or more nucleotides in length). Primers as listed in table 7 are particularly useful for generating PCR products of the following genes: genes encoding the dopamine transporter, dopamine receptor, tryptophan hydroxylase, 5-hydroxytryptamine transporter, 5-hydroxytryptamine receptor, and COMT. When a pair of oligonucleotide primers is used in the same PCR reaction, one primer containing a nucleotide sequence from the coding strand of the nucleic acid and the other primer containing a nucleotide sequence from the non-coding strand of the nucleic acid, a specific region of mammalian DNA can be amplified (i.e., replicated so that multiple correct copies are made). The "coding strand" of a nucleic acid is the non-transcribed strand, which has the same nucleotide sequence as the particular RNA transcript (except that the RNA transcript contains uracil instead of thymidine residues), while the "non-coding strand" of a nucleic acid is the strand that serves as the template for transcription.

A single PCR reaction mixture may contain a pair of oligonucleotide primers. Alternatively, a single reaction mixture may contain multiple oligonucleotide primer pairs, in which case multiple PCR products (e.g., 5, 10, 15, or 20 primer pairs) may be produced. Each primer pair can amplify, for example, one exon or a portion of one exon. Intron sequences may also be amplified.

The exons or introns of the target gene can be amplified and then directly sequenced. Dye primer sequencing can be used to increase the accuracy of detecting heterozygous samples. Alternatively, one or more of the techniques described below may be used to determine the genotype.

For example, allele-specific hybridization can be used to detect sequence variants, including intact haplotypes of mammals. See Stoneking et al, 1991,Am.J.Hum.Genet.48: 370-; and Prince et al, 2001,Genome Res.，11(1): 152-162. Indeed, a sample of DNA or RNA from one or more mammals may be amplified using several pairs of primers and the resulting amplification products may be immobilized on a substrate (e.g., in discrete regions). Hybridization conditions are selected such that the nucleic acid probe can specifically bind to a target sequence, e.g., a variant nucleic acid sequence. Since some sequence variants include only a single nucleotide difference, such hybridization is typically performed under high stringency. High stringency conditions can include washing with low ionic strength solutions and high temperatures. For example, a nucleic acid molecule can be hybridized in 2XSSC (0.3M NaCl/0.03M sodium citrate/0.1% Sodium Dodecyl Sulfate (SDS) at 42 ℃ and washed in 0.1XSSC (0.015M NaCl/0.0015M sodium citrate), 0.1% SDS at 65 ℃.

Allele-specific restriction digests can be performed in the following manner. For nucleotide sequence variants that introduce restriction sites, alleles can be distinguished by restriction digestion with specific restriction enzymes. For sequence variants that do not alter the commonly used restriction sites, mutagenic primers can be designed to introduce restriction sites when variant alleles are present or when wild-type alleles are present. A portion of the target nucleic acid can be amplified using the mutagenic primer and the wild-type primer, followed by digestion with an appropriate restriction endonuclease.

Some variants, such as insertions or deletions of one or more nucleotides, alter the size of the DNA fragment surrounding the variant. Nucleotide insertions or deletions can be assessed by amplifying the region surrounding the variant and determining the size of the amplified product compared to a size standard. For example, a region of the target gene may be amplified using primers placed from either side of the variant. One of the primers is typically labeled with, for example, a fluorescent moiety to facilitate size resolution. The amplification product can be electrophoresed through an acrylamide gel with a set of size standards labeled with a fluorescent moiety different from the primer.

PCR conditions and primers can be developed such that the product is amplified only in the presence of variant alleles or only in the presence of wild-type alleles (MSPCR or allele-specific PCR). For example, patient DNA and control can be amplified separately using wild-type primers or variant allele-specific primers. Each set of reactions was then checked for the presence of amplified product using standard methods to visualize DNA. For example, the reaction may be run through an agarose gel, stained with ethidium bromide or other DNA intercalating dyes to render the DNA visible. In the DNA samples of heterozygous patients, reaction products were detected in each reaction. Patient samples containing only the wild-type allele had amplification products only in the reaction using the wild-type primer. Similarly, patient samples containing only variant alleles have amplification products only in reactions using variant primers. Allele-specific PCR can also be performed using allele-specific primers that incorporate two universal energy transfer labeled primers (e.g., one labeled with a green dye such as fluorescein, and one labeled with a red dye such as sulforhodamine)Label) is added. The amplification products can be analyzed for green and red fluorescence in a plate reader. See, e.g., Myakishev et al, 2001,Genome 11(1)：163-169。

the mismatch cleavage method can also be used to detect different sequences, amplified by PCR, subsequently hybridized to the wild type sequence and cleaved at the point of mismatch. Chemical reagents such as carbodiimide or hydroxylamine and osmium tetroxide can be used to modify mismatched nucleotides to facilitate cleavage.

Kits for detecting a number of cytochrome P450 variants are also available on the market. For example, the TAG-ITTM kit is available from Tm Biosciences Corporation (Toronto, Ontario).

Selecting therapeutic agents

After determining the genotype for each gene in the panel, therapeutic agents can be selected. Typically, the selection involves correlating the genotype of the cytochrome P450 genes with the ability of each cytochrome P450 enzyme encoded by each cytochrome P450 gene to metabolize the therapeutic drug. The genotypes of other genes of interest in the panel (e.g., the 5-hydroxytryptamine transporter and the 5-hydroxytryptamine receptor 2A) can be correlated with the ability of the patient to respond to the therapeutic agent.

Algorithms can be used to select the most appropriate therapeutic drug for an individual patient. The design algorithm requires initial identification of the phenotype, which provides an initial identification of the range of possible therapeutic agents. In the next step of the algorithm, the results of the analysis of the target genes may be sequentially input. The possible list of appropriate therapeutic drugs may then be ranked (rank-ordered) in an algorithmic equation based on the particular cofactor, which assigns a positive, negative, or neutral probability score to each identified therapeutic drug in the initially identified possible selected group. This process adjusts the rank ordering based on the genotype polymorphisms carried by the patient. The result of the CYP gene analysis can be introduced at the next entry point (entrypoint) in the algorithm equation. The algorithmic analysis was designed to place the therapeutic drugs into 3 categories: 1) use of an acceptable therapeutic agent that has a high likelihood of normal metabolism in individuals with a particular genotype, 2) a cautious use of a therapeutic agent (e.g., a therapeutic agent may require some adjustment of administration based on atypical metabolism); and 3) therapeutic agents that should be avoided or that should be carefully and monitored during use, for example due to possible difficulties in administration. At this point in the selection process, data regarding the therapeutic drug response of the patient's first and second relatives may be entered into the algorithmic equations that relate the therapeutic drug selection for the drugs in the first list that have been identified. After adjusting the rank ordering, appropriate therapeutic drugs can be calculated based on the clinical response of the family members.

The selection of an appropriate therapeutic agent may be further enhanced by the inclusion of target data and drug metabolism-related data. This can determine the effect of CYP products on the clinical response of a particular patient. Including, for example, target data and drug metabolism-related data, provides information on the amount of drug available, the ability of the patient to use the drug, and the quality of the target for the drug receptor, providing a reasonable method of selecting a therapeutic drug.

An example of such a method is the selection of an appropriate antidepressant for a given patient. After the phenotype of depression is established, the initial range of antidepressant therapeutic drugs is identified. For example, citalopram, fluvoxamine, bupropion, escitalopram, sertraline, mirtazapine, fluoxetine, venlafaxine, amitriptyline, imipramine, paroxetine and nortriptyline may be the initial range of antidepressant drug therapies. It will be appreciated that the initial range of therapeutic drugs may vary as other antidepressants become available or other pharmacogenomic data are available. Data for genotyping of interest is then entered. If the genotype reflection is revealed in the first component of the algorithm to be weak for 2D6 metabolism and normal for 1A2 metabolism, the ranking of the therapeutic agents would identify fluvoxamine and bupropion (buproprion) as being potentially both effective and manageable at the recommended dose. At the next stage of the algorithm, if the individual is homozygous for the long form of the 5-hydroxytryptamine transporter gene, it is confirmed that fluvoxamine would be acceptable for use. The output values of the algorithm may then be integrated with the historical data. For example, if a family member has responded well to fluvoxamine, this will confirm that the therapeutic is acceptable for use, or if a first or second relative has a problem responding to the therapeutic, an alternative may be made.

Computer system

The techniques described herein may be implemented in a computer system having a processor executing specific instructions in a computer program. The computer system may be arranged to output a therapeutic drug calendar based on the genotype of the receiving patient. In particular, the computer program may include instructions that instruct the system to select the most appropriate therapeutic drug (e.g., psychotherapeutic drug) for an individual patient.

The following are examples of features that may be included in a system. The computer program can be configured to enable a computer system to identify a phenotype based on the received data and provide a preliminary identification of a range of possible therapeutic drugs. The system may be capable of ranking the identified therapeutic drug classes based on the particular cofactor in the algorithm equation. The system may be capable of adjusting the rank ordering based on the genotype polymorphisms carried by the patient. The system may be able to adjust the rank ordering based on clinical responses of family members such as patients.

FIG. 1 is a block diagram of a computer system 100 that may be used in the above-described operations, according to one implementation. The system 100 includes a processor 110, a memory 120, a storage device 130, and an input/output device 140. Each of the components 110, 120, 130, and 140 are interconnected using a system bus 150. The system may include an analysis device 160 to determine the patient's genotype.

The processor 110 is capable of processing instructions for execution in the system 100. In one embodiment, the processor 110 is a single-threaded processor. In another implementation, the processor 110 is a multi-threaded processor. The processor 110 is capable of processing instructions stored in the memory 120 or the storage device 130, including receiving or transmitting information by the input/output device 140.

The memory 120 stores information within the system 100. In one implementation, the memory 120 is a computer-readable medium. In one embodiment, memory 120 is a volatile memory unit. In another implementation, the memory 120 is a non-volatile memory unit.

The storage device 130 is capable of providing mass storage for the system 100. In one embodiment, storage device 130 is a computer-readable medium. In various different embodiments, storage device 130 may be a floppy disk device, a hard disk device, an optical disk device, or a tape device.

The input/output device 140 provides input/output operations for the system 100. In one embodiment, the input/output device 140 includes a keyboard and/or pointing device. In one embodiment, the input/output device 140 includes a presentation unit for presenting a graphical user interface.

The system 100 may be used to build a database. Fig. 2 shows a flow chart of a method 200 of constructing a database for selecting a therapeutic drug for a patient. Preferably, the method 200 is performed in the system 100. For example, the computer program product may include instructions that cause the processor 110 to perform the steps of the method 200. The method 200 includes the following steps.

At step 210, a plurality of genotypes 170 of a set of genes is received. The computer program in system 100 may include instructions that cause the input/output device 140 to present a suitable graphical user interface that may prompt the user to enter the genotype 170 using the input/output device 140, such as a keyboard.

At step 220, a plurality of treatment medications 180 is received. The treatment history 180 is assigned based on the genotype 170. The user may enter the medication calendar 180 using an input/output device 140, such as a keyboard. For example, the treatment medications calendar 180 may include information 190 about at least one treatment medication.

At step 230, the received genotypes 170 and therapeutic agent histories 180 are stored such that each therapeutic agent history 180 is associated with one of the genotypes 170. The system 100 may store the treatment history 180 and the genotype 170 in the storage device 130. For example, upon completion of storage, the system 100 may identify a particular one of the therapeutic agents 180 associated with a particular genotype 170. After identifying the treatment medication calendar 180, the system 100 may obtain information 190 contained in the identified treatment medication calendar 180, as described in the examples below.

The system 100 may be used to select a therapeutic drug. Fig. 3 shows a flow chart of a method 300 of selecting a therapeutic agent for a patient. Preferably, the method 300 is performed in the system 100. For example, the computer program product may include instructions that cause the processor 110 to perform the steps of the method 300. The method 300 includes the following steps.

At step 310, the genotypes of a set of genes of a patient are received. The user may input the genotype via the input/output device 140. For example, a user may use the analysis device 160 (which may or may not be connected to the system 100) to obtain a genotype for a set of genes of a patient. The user may enter the patient's genotype on an input/output device 140, such as a keyboard, to be received by the system 100.

The genotype may be received directly from the analysis device 160. For example, the analysis device 160 may include a processor and suitable software to enable it to communicate over a network. The system 100 may be connected to the analysis device 160 by an input/output device 140, such as a network adapter, and receive the patient's genotype directly.

At step 320, a therapeutic agent associated with the patient's genotype is identified 180. For example, system 100 may conduct a database search in storage device 130. Specifically, the system 100 may acquire the genotype 170 for an individual treatment session 180 until a match is found. Optional step 325 will be described below.

Step 330, the identified therapeutic agent is output 180 in response to receiving the patient's genotype. The system may output the identified medication history 180 by the input/output device 140. For example, the identified treatment history may be printed or presented in a suitable graphical user interface on a display device. As another example, the system 100 may transmit the identified medication history over a network, such as a local area network or the Internet to which the input/output device 140 is connected.

The treatment medications calendar 180 may be created with flexibility in the manner in which they are output by the system 100. For example, the information 190 in the one or more therapeutic drugs calendar 180 may include a ranking of a plurality of therapeutic drugs. The program may include instructions to apply rules to the received patient's genotype and adjust the ranking accordingly. In such implementations, the method 300 may include an optional step 325 of adjusting the ranking prior to outputting the identified treatment history. For example, the system 100 can receive the patient-borne genotype polymorphism (optionally in the same manner that the patient's genotype is received) and adjust the ranking accordingly in step 325. As another example, step 325 may include adjusting the ranking based on clinical response. The clinical response may be received by the system 100 in the same manner as the patient's genotype is received. The ranking may be adjusted based on the clinical response of the patient's family members, for example.

The treatment medications calendar 180 may be updated as needed. For example, the introduction of new therapeutic drugs on the market may suggest revising one or more of the existing therapeutic drug histories. New therapeutic drugs may also serve as the basis for creating new therapeutic drug histories. The adjustment or creation of the therapeutic drug calendar may be performed substantially as described above.

The therapeutic agent calendar 180 may be used for therapeutic agent selection in the same system in which it was created or in a different system. That is, the system 100 may first be used to build a database of therapeutic drug histories 180, and then the system 100 may be used to select a therapeutic drug calendar for a particular patient genotype. As another example, in accordance with the present invention, one or more therapeutic drugs 180 may be transmitted for remote processing in a computer readable medium, such as a global computer network.

Article of manufacture

In one embodiment, the article of manufacture is a composition comprising an oligonucleotide primer. Typically such compositions will comprise a first oligonucleotide primer and a second oligonucleotide primer, each 10 to 50 nucleotides in length, which can be subjected to PCR conditions in combination with mammalian genomic DNA to produce a nucleic acid product corresponding to a region of interest within a target gene. The composition may also contain buffers and other reagents required for PCR (e.g., DNA polymerase or nucleotides). Furthermore, the composition may comprise one or more additional pairs of oligonucleotide primers (e.g., 5, 10, 15, or 20 primer pairs) such that a variety of nucleic acid products may be produced.

In other embodiments, an article of manufacture comprises a population of nucleic acid molecules immobilized on a substrate. Suitable substrates provide the basis for nucleic acid immobilization, and in some embodiments allow for immobilization of nucleic acids to discrete regions. In embodiments where the substrate comprises a plurality of discrete regions, different populations of isolated nucleic acids may be immobilized to each discrete region. The different populations of nucleic acid molecules independently can include nucleic acid molecules for detecting one or more of the alleles listed in tables 1 and 2.

Suitable substrates may be of any shape or form and may be constructed from, for example, glass, silicon, metal, plastic, cellulose, or composite materials. For example, suitable substrates may include multiwell plates or membranes, slides, chips, or polystyrene or magnetic beads. The nucleic acid molecule or polypeptide may be synthesized in situ, immobilized directly to the substrate, or immobilized by a linker, including by covalent, ionic, or physical bonds. Linkers for immobilizing nucleic acids and polypeptides include reversible or cleavable linkers, as known in the art. See, for example, U.S. Pat. No.5,451,683 and WO 98/20019. The immobilized nucleic acid molecule is typically about 20 nucleotides in length, but can vary in length from about 10 nucleotides to about 1000 or more nucleotides.

The invention will be further described by the following examples, which do not limit the scope of the invention described by the claims.

Examples

Example 1

CYP450 genotype of patient

This test highlights the results of an assessment of adolescents who have an atypical response to selective 5-hydroxytryptamine reuptake inhibitors. The following deficient CYP2D6 alleles were detected: 2(1661G > C and 2850C > T),. 3(2549A > del),. 4(1661G > C and 1846G > a),. 10(100C > T, 1661G > C and 1846G),. 17(1023C > T and 2850C > T). The absence of CYP2D6(× 5del) was detected by electrophoresis after PCR amplification. Tandem repeats of the CYP2D6 gene are detected by amplifying repeat-specific sequences and detected by subsequent electrophoresis.

Genomic DNA was amplified by polymerase chain reaction in a practical biological systems Thermal Cycler (applied biosystems Thermal Cycler) using the following primers. In the following examples, the letters "f" and "r" indicate the direction of synthesis ("f" indicates forward/sense synthesis reaction and "r" indicates reverse/antisense synthesis reaction). The amplified DNA was hybridized with oligonucleotide probes specific for normal and polymorphic variants (described below) and detected using the Nanogen molecular biology workstation. The oligonucleotide probes are fluorescently labeled (e.g., CY3 or CY 5).

Pre-nested (PreNest) oligonucleotides(initial amplification)

An amplification primer:

CYP2D6 pre-nested B F5'-CCA GAA GGC TTT GCA GGC TTC A-3' (bases 1281-1302, SEQ ID NO: 1)

CYP2D6 pre-nested B R5'-ACT GAG CCC TGG GAG GTA GGT A-3' (bases 6357-6378, SEQ ID NO: 2)

Nested oligonucleotides (second amplification)

CYP2D6(C100T)

An amplification primer:

^■CYP2D6(100) B f 5 'GGC CTA CCC TGG GTA AGG GCC TGG AGCAGG A-3' (base 1579-1594, SEQ ID NO: 3)

^■CYP2D6(100) B r 5 '-/5 Bio/CCT GGT CGA AGC AGT AT-3' (bases 2394-

Probes and Stabilizers (Stabilizers):

CYP2D6C100T wild type 5 '-/5 Cy3/GCA CGC TAC C-3' (base 1700-

CYP2D6C100T polymorphism 5 '-/5 Cy5/GCA CGC TAC T-3' (base 1700-

CYP2D6C100T stabilizer 5 'CAC CAG GCC CCC TGC CAC TG-3' (base 1711-1731, SEQ ID NO: 7)

CYP2D6(C1023T)

An amplification primer:

^■CYP2D6(1023) B f 5'-CCT GCT CAC TCC TGG TAG CC-3' (base 2394-

■ CYP2D6(1023) B r 5 '-/5 Bio/CTG TTT CAT GTC CAC GAC C-3' (bases 2760-

Probe and stabilizer:

CYP2D6(1023) wild type 5 '-/5 Cy3/TGC CCA TCA C-3' (bases 2632-

CYP2D6(1023) polymorphism 5 '-/5 Cy3/TGC CCA TCA T-3' (base 2632-

CYP2D6(1023) stabilizer 5 'CCA GAT CCT GGG TTT CGG GCC GCG-3' (base 2643-2667, SEQ ID NO: 12)

CYP2D6(G1661C and G1846A)

Note that: the amplicon has 2 polymorphisms.

An amplification primer:

^■CYP2D6(1661, 1846) B f 5'-CAG AGG CGC TTC TCC G-3' (bases 3266-3282, SEQ ID NO: 13)

^■CYP2D6(1661, 1846) B r 5 '-/5 Bio/CTC GGT CTC TCG CTC CGC AC-3' (bases 3630-3650, SEQ ID NO: 14)

Probe and stabilizer:

CYP2D6(1661) wild type 5 '-/5 Cy3/CTT CTC CGT G-3' (bases 3266-3276, SEQ ID NO: 15)

CYP2D6(1661) polymorphism 5 '-/5 Cy3/CTT CTC CGT C-3' (bases 3266-3276, SEQ ID NO: 16)

CYP2D6(1661) stabilizer 5'-TCC ACC TTG CGC AAC TTG GGC CTGGG-3' (base 3277-3303, SEQ ID NO: 17)

CYP2D6(1846) wild type 5 '-/5 Cy3/CCA CCC CCA G-3' (bases 3460-

CYP2D6(1846) polymorphism 5 '-/5 Cy3/CCA CCC CCA A-3' (base 3460-

CYP2D6(1846) stabilizer 5'-GAC GCC CCT TTC GCC CCA ACG-3' (base 3471-3492, SEQ ID NO: 20)

CYP2D6(A2549del and C2850T)

An amplification primer:

CY P2D6(A2549del and C2850T) B f 5'-TGA GAC TTG TCC AGG TGAAC-3' (bases 4001-

^■CYP2D6(A2549del and C2850T) B r 5 '-/5 Bio/CCC AGA TGG GCT CACGCTGC-3' (bases 4558-4578, SEQ ID NO: 22)

Probe and stabilizer:

CYP2D6(2549) wild type 5 '-/5 Cy3/ACT GAG CAC A-3' (base 4161-4171, SEQ ID NO: 23)

CYP2D6(2549) polymorphism 5 '-/5 Cy3/ACT GAG CAC G-3' (base 4161-4171, SEQ ID NO: 24)

CYP2D6(2549) stabilizer 5 '-GGA TGA CCT GGG ACC CAG CCC AGCC 3' (base 4172-4199, SEQ ID NO: 25)

CYP2D6(2850) wild type 5 '-/5 Cy3/GAG AAC CTG C-3' (bases 4462-4472, SEQ ID NO: 26)

CYP2D6(2850) polymorphism 5 '-/5 Cy3/GAG AAC CTG T-3' (base 4462-4472, SEQ ID NO: 27)

CYP2D6(2850) stabilizer 5'-GCATAG TGG TGG CTG ACC TGT TCTCTG-3' (bases 4473-4500, SEQ ID NO: 28)

PCR amplification

A. Pre-nested (initial amplification):

initial pre-nested PCR for CYP2D6(C1661G), CYP2D6(G1846A), CYP2D6(a2549Del), CYP2D6(C100T), CYP2D6(C2850T), and CYP2D6(C1023T) polymorphisms was performed with the following reagents per sample: mu.L of 25. mu.M pre-nested B F, 0.6. mu.L of 25. mu.M pre-nested B R, 2.5. mu.L of 10 mM dNTP, 39.55. mu.L dH2O, 5.0. mu.L of extension (Expand) buffer 3(Roche), 0.75. mu.L of the extension enzyme mixture (Roche), and 1.0. mu.L of genomic DNA in a total volume of 50. mu.L. PCR was performed as follows: hold at 94 ℃ for 2 minutes, then 30 cycles: denaturation at 94 ℃ for 30 seconds, annealing at 67 ℃ for 30 seconds and extension at 68 ℃ for 7 minutes, followed by final extension at 72 ℃ for 10 minutes. The expected PCR product was ≈ 4.8-5 Kb.

All PCR thereafter was performed using the pre-nested B product as amplification template.

CYP2D6(100)B

(C100T) nested PCR was performed using 1. mu.l of the initial pre-nested B amplicon as template. The PCR product was used to detect CYP2D6(C100T) mutations and the following reagents (25 μ L total volume) were used: o μ l25 μ M CYP2D6(100) B f, 1.0 μ l25 μ M CYP2D6(100) B r, 12.5 μ l Qiagen Hot Start Master Mix, and 9.5 μ l dH₂And O. PCR was performed as follows: held at 95 ℃ for 15 minutes, then 30 cycles: denaturation at 95 ℃ for 30 seconds, annealing at 57 ℃ for 30 seconds and extension at 72 ℃ for 1 minute, followed by final extension at 72 ℃ for 7 minutes.

CYP2D6(1023)B

(C1023T) nested PCR was performed using 1. mu.l of pre-nested B product as template. The PCR product was used to detect CYP2D6(C1023T) mutations and the following reagents (25 μ L total volume) were used: 1.0. mu.l 25. mu.M CYP2D6(1023) B f, 1.0. mu.l 25. mu.M CYP2D6(1023) B r, 12.5. mu.l AmplitaqG01D Master Mix, and 9.5. mu.l dH 2O. PCR was performed as follows: held at 95 ℃ for 10 minutes, then 30 cycles: denaturation at 95 ℃ for 30 seconds, annealing at 59 ℃ for 30 seconds and extension at 72 ℃ for 1 minute, followed by final extension at 72 ℃ for 7 minutes.

CYP2D6(C1661G and G1846A) B

(C166lG and G1846A) nested PCR was performed using 1. mu.l of the pre-nested B product as template. The PCR products were used to detect CYP2D6(C1661G) and CYP2D6(G1846A) mutations and the following reagents (25 μ L total volume) were used: 1.0. mu.l 25. mu.M CYP2D6(1661, 1846) B f, 1.0. mu.l 25. mu.M CYP2D6(1661, 1846) B r, 12.5. mu.l 2X Amplitaq Gold Master Mix and 9.5. mu. ldH 2O. PCR was performed as follows: held at 95 ℃ for 10 minutes, then 30 cycles: denaturation at 95 ℃ for 30 seconds, annealing at 63 ℃ for 30 seconds and extension at 72 ℃ for 1 minute, followed by final extension at 72 ℃ for 7 minutes.

CYP2D6(A2549Del and CYP2D6C2850T) B

(A2549Del and C2850T) nested PCR was performed using 1. mu.l of pre-nested B product as template. The PCR products were used to detect CYP2D6(a2549Del) and CYP2D6(C2850T) mutations and the following reagents (25 μ L total volume) were used: 1.0 μ l25 μ M CYP2D6(2549, 2850) Bf, 1.0 μ l25 μ M CYP2D6(2549, 2850) Br, 12.5 μ l 2X Amplitaq Gold Master Mix, and 9.5 μ ldH 2O. PCR was performed as follows: held at 95 ℃ for 10 minutes, then 30 cycles: denaturation at 95 ℃ for 30 seconds, annealing at 57 ℃ for 30 seconds and extension at 72 ℃ for 1 minute, followed by final extension at 72 ℃ for 10 minutes.

Polymorphisms were detected using the Nanogen molecular biology workstation according to the Nanogen user manual. Samples were desalted on Millipore desalting plates. To a 1.5ml centrifuge tube were added the following reagents: mu.l wild-type probe, 2.0. mu.l polymorphism probe, 2.0. mu.l stabilizer and 44. mu.l high-salt buffer. Each assay has probes and stabilizers that are labeled and specific for it. Vortex the reagents, spin briefly and dispense on the array of nanochips for 5 minutes, place in a dark container. The chips were read every two degrees from 24 ℃ to 50 ℃ by trending method (trending method). It was confirmed that the fluorescent probes hybridized at 24 ℃ and all the probes were dehybridized at 50 ℃. If the Relative Fluorescence Units (RFU) at 50 ℃ on any chip is higher than 80, the chip is immersed (stripeped) in 200. mu.l of 1.0M NaOH for 10 minutes with dH₂Wash 3 times with O and 1 time with high salt buffer. Heterozygote controls (known heterozygotes, one for "normalization" or measurement of RFU, the other to confirm the correct trend pattern observed in pre-genotyped heterozygotes) and negative controls (no DNA) were included in each polymorphism. L-histidine was used to subtract background.

Tables 3-5 summarize the total number of participants in different age groups tested using the above procedure and the inferred metabolic phenotype of CYP2D 6. Table 6 provides the specific 2D6 haplotypes observed in patients with antidepressant drug resistant subtypes of depression.

TABLE 3

Total number of participants in each group (all ages; N86)

CYP2D6 inferred metabolic phenotype

TABLE 4

Total number of participants in each group (age: < 19; N ═ 12)

CYP2D6 inferred metabolic phenotype

TABLE 5

Total number of participants in each group (age: > 18; N ═ 74)

CYP2D6 inferred metabolic phenotype

TABLE 6

Antidepressant therapeutic drug tolerance subtypes for depression¹

Subtype #	2D6 haplotype	2D6 enzymatic action
			100MR	*1/*2P Hz*2	I
101MR	Hz*4	I
			102MR	*1/*10H2*10	I
103MR	Hz*4	I
			104MR	2/' 2P Homozyg 2	I-P
105MR	Hz*11	I
			106MR	Hz*4	I
107MR	Homozygous 3	Is free of
			108MR	Hz*4	I
109MR	2/' 2 homozygous 2	I-P
			110MR	Hz*2	I
111MR	Hz*2	I
			112MR	WT	N
113MR	Hz*2	I
			114MR	WT	N
115MR	WT	N
			116MR	3/'4 complex Hz 3/' 4	P
117MR	Hz*4	I

Subtype #	2D6 haplotype	2D6 enzymatic action
			118MR	Hz*2	I
119MR	Composite Hz 2/' 4	P
			120MR	Hz*4	I
121MR	Hz*4	I
			122MR	Hz*4	I
123MR	WT	N
			124MR	Hz*12	I
125MR	Hz*4	I
			126MR	Hz*4	I
128MR	WT	N
			129MR	Hz*4	I
130MR	Hz*1E	N
			131MR	Homozygous 4	P
132MR	Hz*4	I
			133MR	Hz*2	I
134MR	Composite Hz 2/' 3	P
			135MR	Composite Hz 3/4	P
136MR	Hz*	I
			137MR	Hz*2	I
138MR	Composite Hz 2/' 4	P
			139MR	Composite Hz 2/' 5	P
141MR	Homozygous 5	Is free of
			142MR	Composite Hz 4/' 15	P
143MR	Hz*4	I
			144MR	Hz 17 or 34	I
145MR	Hz*3	I
			146MR	Composite Hz 4/' 5	P
147MR	WT	N
			148MR	Hz*2	I
149MR	WT	N
			150MR	Hz*2	I
155MR	WT	N
			156MR	Hz*4	I
157MR	Hz*2	I
			158MR	Hz*4	I
159MR	Hz*2	I
			160MR	WT	N
161MR	Hz*2	I
			162MR	WT	N
164MR	Hz*2	I

Subtype #	2D6 haplotype	2D6 enzymatic action
			165MR	Hz*2	I
166MR	WT	N
			167MR	Composite Hz 2/' 4	P
168MR	Composite Hz 2/' 4	P
			169MR	WT	N
170MR	Hz*2	I
			171MR	Composite Hz 2/' 3	P
172MR	Hz 2 w/repetition	Incomplete action
			173MR	Hz*2	I
174MR	Repetition of Hz 4 w/balance	N
			175MR	Hz*2	I
176MR	Hz*4	I
			177MR	Hz*2	I
179MR	Homozygous 2	I-P
			180MR	Homozygous 2	I-P
181MR	WT	N
			182MR	Hz*4	I
183MR	WT	N
			184MR	Hz*2	I
185MR	WT	N
			187MR	Composite Hz 2/' 4	P
188MR	Hz*4	I
			189MR	Hz*10	I
190MR	WT	N
			191MR	Hz*2	I
192MR	Hz*4	I
			193MR	Hz*2	I
194MR	Hz*3	I
			195MR	Hz 17 or 34	I
196MR	Hz*4	I
			197MR	Composite Hz 2/' 4	P
198MR	Hz*2	I
			199MR	Composite Hz 2/' 4	P

¹P is weak; n is normal; i-medium, HI-higher inducibility; WT ═ wild type

All of these adolescents were found to have an atypical haplotype of 2D6 (see tables 3-5). Our initial results of analyzing the cytochrome P450 gene alleles in adult human samples also demonstrated the high variability of the polymorphism of the 2D6 gene (see tables 3-5). It was also noted that adolescents with polymorphisms weakly associated with 2D6 metabolism had a clinical history characterized by poor response or high frequency of side effects to therapeutic drugs when they had been treated with a selective 5-hydroxytryptamine reuptake inhibitor, which is metabolized by the 2D6 enzyme.

Example 2

Determination of dopamine transporters, dopamine receptors,Tryptophan hydroxylase, Of the 5-hydroxytryptamine receptor and COMT genesGenotype(s)

The following probes and primers of Table 7 can be generated to detect the genotypes of the dopamine transporter (DAT1, SLC6A3), dopamine receptor (DRD1, DRD2, DRD3, DRD4, and DRD5), tryptophan hydroxylase (TPH), 5-hydroxytryptamine transporter (5-HTT), 5-hydroxytryptamine receptor (HTR1A, HTR1B, HTR1D, HTR2A, and HTR2C), and COMT genes.

Example 3

The genotypes of the CYP2D6, CYP2C19, and 5HTTR genes were determined in 97 patients using the method described above. Short/long versions of CYP2D6 x 2, x 3, x 4, x 10, x 17 and x 5del alleles, CYP2C19 x 2(a, B), ' 3, ' 4, ' 5(a, B), ' 6, ' 7 and x 8 alleles and the 5HTTR gene were evaluated as indicated in example 1.

Each patient that is genotyped exhibits side effects that are poorly responsive or intolerant to at least one antidepressant therapeutic drug. Based on the genotype information, 18 treatment drug histories were generated (table 8). In table 8, PM is a weak metabolic form; IM is intermediate metabolic type; EM is a broad metabolic type; s is a short form of the gene; l is the long form of the gene. Each treatment regimen provides for the use of acceptable antidepressant treatment medications, antidepressant treatment medications that should be used with caution, and antidepressant treatment medications that should be avoided or used with caution and closely monitored.

TABLE 8

Therapeutic drug algorithm microarray results for pharmacological studies

Genotype catalogue of key genes	2D6	2C19	5HTTTR	Genotype frequency in pharmacological studies
					Calendar of therapeutic drugs 1	PM	PM	s/s	2
Calendar of therapeutic drugs 2	PM	PM	s/l	2
					Therapeutic agentsCalendar 3	PM	PM	l/l	1
Calendar of therapeutic drugs 4	PM	EM	s/s	1
					Calendar 5 of therapeutic drugs	PM	EM	s/l	6
Calendar 6 of therapeutic drugs	PM	EM	l/l	1
					Calendar 7 of therapeutic drugs	IM	PM	s/s	1
Calendar of therapeutic drugs 8	IM	PM	s/l	7
					Medicine for curing diabetesMedicine calendar 9	IM	PM	l/l	3
Calendar 10 of therapeutic drugs	EM	PM	s/s	0
					Calendar 11 of therapeutic drugs	EM	PM	s/l	6
Calendar 12 of therapeutic drugs	EM	PM	l/l	3
					Calendar 13 of therapeutic drugs	IM	EM	s/s	2
Calendar 14 of therapeutic drugs	IM	EM	s/l	17
					Calendar 15 of therapeutic drugs	IM	EM	l/l	16
Calendar 16 of therapeutic drugs	EM	EM	s/s	7
					Calendar of therapeutic drugs 17	EM	EM	s/l	10
Calendar 18 of therapeutic drugs	EM	EM	l/l	12

The therapeutic drug has been associated with 2D6 and 2C19 weak metabotropic patterns and the s/s form of the 5HTTR gene in 1. Bupropion is acceptable for use in patients with the therapeutic agent calendar 1, mirtazapine and fluvoxamine are used with caution, and citalopram, escitalopram, paroxetine, fluoxetine, venlafaxine, amitriptyline, imipramine, nortriptyline and sertraline should be avoided or monitored carefully during use.

Therapeutic agents 2 are associated with the weak metabotropic forms of 2D6 and 2C19 and the s/l form of the 5HTTR gene. Fluvoxamine and bupropion are acceptable for use in patients with treatment course 2, sertraline and mirtazapine are used with caution, and citalopram, escitalopram, paroxetine, fluoxetine, venlafaxine, amitriptyline, imipramine and nortriptyline should be avoided or monitored carefully during use.

Therapeutic drugs are associated with 2D6 and 2C19 weak metabotropic and l/l forms of the 5HTTR gene. Fluvoxamine and bupropion are acceptable for use in patients with the treatment medications calendar 3, sertraline and mirtazapine can be used with caution and close monitoring when citalopram, escitalopram, paroxetine, fluoxetine, venlafaxine, amitriptyline, imipramine and nortriptyline are avoided or used.

Therapeutic agents are associated with the weak 2D6 metabotropic form, the extensive 2C19 metabotropic form and the s/s form of the 5HTTR gene. Bupropion is acceptable for use in patients with the treatment course 4, mirtazapine, citalopram, fluvoxamine and escitalopram are used with caution and close monitoring of the use or avoidance of paroxetine, fluoxetine, venlafaxine, amitriptyline, imipramine, nortriptyline and sertraline.

The therapeutic drug is associated with 2D6 weak metabotropic, 2C19 extensive metabotropic and s/s form of the 5HTTR gene. In patients with the treatment course 5, citalopram, fluvoxamine, bupropion and escitalopram are acceptable for use, sertraline and mirtazapine are used with caution, and paroxetine, fluoxetine, venlafaxine, amitriptyline, imipramine and nortriptyline should be avoided or monitored carefully during use.

The therapeutic agents were associated with 2D6 weak metabotropic, 2C19 extensive metabotropic, and the l/l form of the 5HTTR gene at 6. In patients with the treatment course 6, citalopram, fluvoxamine, bupropion and escitalopram are acceptable for use, sertraline and mirtazapine are used with caution, and paroxetine, fluoxetine, venlafaxine, amitriptyline, imipramine and nortriptyline should be avoided or monitored carefully during use.

Therapeutic agents are associated with the intermediate metabotropic form of 2D6, the weak metabotropic form of 2C19, and the s/s form of the 5HTTR gene. Bupropion is acceptable for use in patients with the treatment medications of calendar 7, mirtazapine, venlafaxine, amitriptyline, nortriptyline, fluvoxamine and sertraline can be used with caution, and citalopram, escitalopram, fluoxetine, imipramine and paroxetine should be avoided or monitored carefully when used.

The therapeutic agents were associated with the intermediate metabotropic form of 2D6, the weak metabotropic form of 2C19, and the s/l form of the 5HTTR gene. Fluvoxamine, bupropion and sertraline are acceptable for use in patients with the therapeutic drug course 8, mirtazapine, paroxetine, venlafaxine, amitriptyline and nortriptyline can be used with caution and close monitoring of the use or avoidance of citalopram, escitalopram, fluoxetine and imipramine.

The therapeutic agents were associated with the intermediate metabotropic form of 2D6, the weak metabotropic form of 2C19, and the l/l form of the 5HTTR gene. Fluvoxamine, bupropion and sertraline are acceptable for use in patients with the therapeutic drug of course 9, mirtazapine, paroxetine, venlafaxine, amitriptyline and nortriptyline can be used with caution and close monitoring of the use or avoidance of citalopram, escitalopram, fluoxetine and imipramine.

Therapeutic agents 10 are associated with the extensive metabotropic form of 2D6, the weak metabotropic form of 2C19, and the s/s form of the 5HTTR gene. Bupropion and venlafaxine are acceptable for use in patients with the therapeutic agent of course 10, mirtazapine, nortriptyline, imipramine, amitriptyline, sertraline, paroxetine and fluoxetine are used with caution and close monitoring of the use or avoidance of fluvoxamine, citalopram and escitalopram.

The therapeutic drug is associated with 2D6 extensive metabotropic, 2C19 weak metabotropic and s/l forms of the 5HTTR gene. In patients with the treatment course 11, sertraline, venlafaxine, paroxetine, bupropion, and fluoxetine are acceptable for use, mirtazapine, fluvoxamine, nortriptyline, citalopram, escitalopram, imipramine, and amitriptyline may be used cautiously.

The therapeutic drug is 12 related to the extensive metabotropic form of 2D6, the weak metabotropic form of 2C19, and the l/l form of the 5HTTR gene. In patients with the treatment course 12, sertraline, venlafaxine, paroxetine, bupropion, and fluoxetine are acceptable for use, mirtazapine, fluvoxamine, nortriptyline, citalopram, escitalopram, imipramine, and amitriptyline may be used cautiously.

The therapeutic agents were associated with the intermediate metabotropic form of 2D6, the extensive metabotropic form of 2C19, and the s/s form of the 5HTTR gene. Bupropion and mirtazapine are acceptable for use in patients with 13 th of therapeutic history, venlafaxine, amitriptyline, imipramine, nortriptyline, citalopram, fluvoxamine, escitalopram and sertraline can be used with caution, and paroxetine and fluoxetine should be avoided or monitored carefully when used.

The therapeutic drug is associated with medium grade 2D6 metabotropic, extensive metabotropic 2C19 and s/l form of the 5HTTR gene. In patients with the treatment course 14, citalopram, fluvoxamine, bupropion, escitalopram, sertraline and mirtazapine are acceptable for use, and paroxetine, fluoxetine, venlafaxine, amitriptyline, imipramine and nortriptyline may be used with caution.

The therapeutic agents were associated with medium grade 2D6 metabotropic, extensive metabotropic 2C19 and the l/l form of the 5HTTR gene. In patients with the therapeutic agent of claim 15, citalopram, fluvoxamine, bupropion, escitalopram, sertraline and mirtazapine are acceptable for use, and paroxetine, fluoxetine, venlafaxine, amitriptyline, imipramine and nortriptyline may be used with caution.

The therapeutic agents were associated 16 with a broad metabolic pattern of 2D6, a broad metabolic pattern of 2C19 and the s/s form of the 5HTTR gene. In patients with the therapeutic drug calendar of 16, bupropion, mirtazapine, venlafaxine, amitriptyline, imipramine and nortriptyline are acceptable for use, and paroxetine, fluoxetine, fluvoxamine, sertraline, escitalopram and citalopram can be used cautiously.

The therapeutic agents were associated with a broad metabolic pattern of 2D6, a broad metabolic pattern of 2C19, and the s/l form of the 5HTTR gene. The therapeutic agents are associated with a broad metabolic pattern of 2D6, a broad metabolic pattern of 2C19 and the l/l form of the 5HTTR gene at 18. In patients with the therapeutic agents of claim 17 or 18, citalopram, fluvoxamine, bupropion, escitalopram, sertraline, mirtazapine, paroxetine, fluoxetine, venlafaxine, amitriptyline, imipramine and nortriptyline are acceptable for use.

Example 4

The genotypes of the CYP2D6, CYP2C19, CYP3a4, CYP1a2 and HTR2A genes were determined in 10 patients using the above method. The genotype of the 5HTTR gene was determined by amplifying the polymorphic region of the gene using primers flanking that region. The size of the reaction product was resolved using electrophoresis (Agilent Technologies). CYP3a4 x 1B, x 2, x 5, x 6, x 12, x 13, x 15A, x 17 and x 18A alleles, CYP1a 2x 1F alleles and HTR2A 102 polymorphisms were tested.

Based on the CYP2D6 genotype, patients were classified as either weak, intermediate, extensive or ultrafast metabolic. For each of the CYP2C19, CYP3a4 and CYP1a2 genotypes, patients were classified as either extensive or non-extensive (i.e., weak) metabolic. For the 5HTTR genotype, patients were classified as having short/short (s/s) form of the gene, short/long (s/l) form of the gene, or long/long (l/l) form of the gene. For the HR2A genotype, patients were classified as having the C/C allele or as having the T/T or T/C allele.

The following 6 rules were used to sort genotypes into 192 treatment recommendations. The first rule relates to the CYP2D6 genotype. For the weakly and ultrafast metabolic types, the CYP2D6 substrate is placed in a "carefully monitored and carefully used" list. For the intermediate metabolic types, the specialized (exclusive) CYP2D6 substrate was placed in a "cautious use to avoid or closely monitor" list. For the intermediate metabolic forms, substrates that are partially metabolized by CYP2D6 are placed in a "cautious" list. For a wide range of metabolic forms, CYP2D6 substrates are placed in a "use acceptable" list.

After the first rule is implemented, a second rule involving the genotype of 2C19 is implemented. For the weakly metabolic forms based on their CYP2C19 genotype, specialized CYP2C19 substrates were placed in a "avoid or closely monitor cautious" list. For broad or intermediate metabolic types based on their CYP2C19 genotype, CYP2C19 substrates were placed in a "use acceptable" list. Substrates metabolized primarily by CYP2D6 and CYP2C19 are placed in a "cautious" list of avoiding or closely monitoring if CYP2C19 is weak and CYP2D6 is medium or weak, or if CYP2C19 is weak and CYP2D6 is extensive.

The first rule and the second rule are followed by a third rule relating to the genotype of 3a 4. For the weakly metabolic forms based on their CYP3a4 genotype, a specialized CYP3a4 substrate was placed in a "avoid or closely monitor cautious" list. For broad or intermediate metabolic types based on their CYP3a4 genotype, CYP3a4 substrates were placed in a "use acceptable" list. Substrates that are metabolized primarily by any two of the following three enzymes: CYP2D6, CYP2C19, or CYP3a4, are placed in a "cautious" list to avoid or closely monitor for very cautious use if any two of the three enzymes are of a medium or weak metabolic type, or a "cautious" list if one of the two most major enzymes is a weak metabolic type.

The fourth rule involving genotype 1a2 was implemented after the first three rules were implemented. For the weakly metabolised forms based on the CYP1a2 genotype, the CYP1a2 substrate was placed on a "cautious use of avoiding or closely monitoring" list (including fluvoxamine as the primary CYP1a2 substrate). For the weak metabolic forms based on the CYP1a2 genotype, substrates with the CYP1a2 pathway as one of their major pathways were placed in a "cautious" list.

After the first four rules are implemented, a fifth rule involving the 5HTTR genotype is implemented. For homozygous short genotypes (s/s) of the 5-hydroxytryptamine transporter gene, SSRIs classified as "acceptable for use" were transferred to the "caution use" list. For the pure truncated genotype (s/s) of the 5-hydroxytryptamine transporter gene, SSRIs classified as "cautious" were transferred to a "avoid or closely monitor cautious" list. Antidepressants other than the SSRI class are not based on the 5HTTR genotype transfer list.

After the first five rules were implemented, a sixth rule involving the HR2A genotype was implemented. For the "mutant form" of the 5-hydroxytryptamine receptor 2A gene, if paroxetine is in the "use acceptable" category, it is transferred to the "caution use" category. If paroxetine is in the "cautious" list, it is transferred to "avoid or closely monitor very cautious use".

Based on the patient's genotype and associated therapeutic history, the appropriate therapeutic is selected for the patient. Table 9 summarizes data for 10 patients. The following presents a case study of 4 out of 7 patients.

The first case was a 16-year-old white female who was genotyped to find a weak CYP2D6 metabolite (. x 3/. x 9) and a weak CYP2C19 metabolite (. x 2/. x 2). She was diagnosed with ADHD at age 6 and failed treatment. She then developed depressive and mood disorder (mood dysregulation) symptoms for treatment with a number of therapeutic drugs metabolized by CYP2D6, such as EFFEXOR. Furthermore, she was homozygous for the short form of the 5-hydroxytryptamine transporter gene. Based on her specific genotypes of these key genes, the algorithm demonstrated a relatively small number of possible strategies to control her depression. The therapeutic drug history generated based on her s/s forms of HR2ATT/TC, 5HTTR, the broad metabolom of 1A2 and 3A4, and the genotype of the weak metabolom of 2C19 and 2D6, indicates that bupropion is acceptable for use, that mirtazapine and fluvoxamine (fluoxamine) should be used with caution, and that citalopram, escitalopram, paroxetine, fluoxetine, venlafaxine, amitriptyline, imipramine, nortriptyline, and sertraline should be avoided or used with caution and monitored.

Before genotyping, she had an especially abnormal side effect on citalopram, as she developed hallucinations (incidence < 1%). This atypical and dangerous side effect could be avoided if she was genotyped before receiving the treatment. The metabolic enzyme primarily involved in citalopram metabolism is CYP2C19 and the second metabolic pathway is CYP2D 6. In this patient, both pathways are non-functional. Thus, even at moderate doses of citalopram, the patient developed an acute auditory and visual hallucination, which was subsequently treated with atypical anti-stress agents. Furthermore, she required three acute hospitalizations and a partial hospitalization for up to 2 months prior to being assigned to the public hospital due to the failure to develop an effective treatment plan for that patient. She was unsuccessfully tested no more than a dozen psychotherapeutic drugs. Of course, if the clinician knows that she cannot take advantage of many medications previously prescribed at their standard doses, her care will be greatly improved and some hospitalizations may be avoided. In addition, her 5-hydroxytryptamine transporter genotype (C/C) would explain the poor response to many of the previously prescribed therapeutic drugs. After genotyping, she was treated with a combination of two mood stabilizers metabolized by alternative pathways (lamital (lamotrigine) and TRILEPTAL (oxcarbazepine)). She also tolerated ABILIFY (aripiprazole) treatment, which is metabolized by CYP3A4 in addition to CYP2D6, for which she had an intrinsic metabolic pathway. She is now reasonably dosed.

The second case was a 42 year old caucasian woman with a history of long-term depression and combined anxiety and years of psychotherapeutic drug intolerance. The patient underwent a comprehensive neurological assessment and failed to explain his symptoms providing quality. This patient has been treated in the past with at least 9 psychotherapeutic drugs. Significant examples of CYP2D6 intolerance include the development of acute hallucinations and very low amitriptyline resistance when paroxetine is taken. The patient was poorly metabolised to CYP2D6 and moderately metabolised to CYP2C 19. This represents a rare genotype, explaining the difficulty of tolerating citalopram treatment. Indeed, when taking citalopram she does have irritability and lethargy. To further complicate the problem, she is also homozygous for the short form of the 5-hydroxytryptamine transporter, which correlates with poor response to 5-hydroxytryptamine reuptake (regenerative) inhibitors. She underwent multiple outpatients for her psychiatric problems. Knowing her genotype early in her history would obviously allow her clinician to avoid many therapeutic drugs she cannot tolerate. The therapeutic drug history based on her s/s forms of HR2A TT/TC, 5HTTR, the broad metabolom of 1A2 and 3A4, and the genotype of the weak metabolom of 2C19 and 2D6 indicated that bupropion is acceptable for use, mirtazapine and fluvoxamine (fluoxamine) should be used with caution, and citalopram, escitalopram, paroxetine, fluoxetine, venlafaxine, amitriptyline, imipramine, nortriptyline, and sertraline should be avoided or used with caution and monitored.

The third case history was a 67 year old female diagnosed with major depressive disorder that had been treated with 7 different therapeutic agents in the past. These failed to be effective and she required two psychiatric hospitalizations. She was identified as CYP2D6 (./9) and CYP2C19 (./2) weakly metabolised forms. Furthermore, she is homozygous for the long form of the 5-hydroxytryptamine transporter gene. Based on her l/l form of HR2A C/C, 5HTTR, the broad metabolic form of 1A2 and 3A4, and the genotype of the weak metabolic form of 2C19 and 2D6, the relevant therapeutic guidelines indicate that fluvoxamine (fluoxamine) and bupropion are acceptable for use, that sertraline and mirtazapine should be used with caution, and that citalopram, escitalopram, paroxetine, fluoxetine, venlafaxine, amitriptyline, imipramine and nortriptyline should be avoided or used with caution and monitored. If her genotype is known, she would likely be expected to have adverse effects on EFFEXOR (venlafaxine) and LEXAPRO (escitalopram oxalate). Since 2 of the 3 metabolic pathways involved in LEXAPRO metabolism are completely inactive, it is likely that this result will be predicted. After her genotyping, she was successfully treated with electroconvulsive therapy (ECT) and at the same time successfully treated with REMERON (mirtazapine), trazodone and KLONOPIN (clonazepam), which have alternative metabolic pathways for CYP2D6 and CYP2C 19.

The fourth disease case is a 50 year old caucasian woman, which has been diagnosed with dysthymia and chronic anxiety. She had been treated with a variety of psychotropic medications, including two CYP2D6 medications that she was unable to tolerate. Her genotypes indicated that she was a weak metabolised form of CYP2D6 (. about.4/. about.4) and a medium metabolised form of CYP2C19 (. about.1/. about.2). She was heterozygous for the long and short forms of the 5-hydroxytryptamine transporter gene. Based on her l/s form of HR2A C/C, 5HTTR, the broad metabolom of 1A2 and 3A4, and the genotype of the weak metabolom of 2C19 and 2D6, the relevant therapeutic guidelines indicate that fluvoxamine (fluoxamine) and bupropion are acceptable for use, that sertraline and mirtazapine should be used with caution, and that citalopram, escitalopram, paroxetine, fluoxetine, venlafaxine, amitriptyline, imipramine and nortriptyline should be avoided or used with caution and monitored.

The patient was hospitalized several times at considerable cost. She also found it difficult to tolerate a range of non-psychotherapeutic drugs, including PROZAC (fluoxetine) and ZOLOFT (sertraline) in only one dose, which found extreme discomfort. Of particular interest, she eventually responded to sub-clinical doses of paroxetine, which represents one example of a method of using an algorithm to find an effective therapeutic drug to administer (that is not tolerated at normal doses).

Based on the l/l form of patient 5 HR2A C/C, 5HTTR, the broad metabolom of 1a2 and 3a4, and the genotype of the weak metabolom of 2C19 and 2D6, the relevant therapeutic regimens indicate that fluvoxamine (fluoxamine) and bupropion are acceptable for use, that sertraline and mirtazapine should be used with caution, and that citalopram, escitalopram, paroxetine, fluoxetine, venlafaxine, amitriptyline, imipramine and nortriptyline should be avoided or used with caution and monitored.

The therapeutic drug history associated with patient genotype 6 (HTR2A CC; 5HTTR l/s; 1A2, 3A4 and 2D6 are extensive; and 2C19 weak) indicates that sertraline, venlafaxine, bupropion and fluoxetine are acceptable for use, and mirtazapine, fluvoxamine, nortriptyline, citalopram, escitalopram, imipramine, amitriptyline and paroxetine may be used with caution.

The therapeutic regiments associated with patient genotype 7 (HTR2A TT/TC; 5HTTR l/s; 1A2, 3A4 and 2C19 are extensive; and 2D6 ultrafast) indicate that citalopram, fluvoxamine, bupropion and escitalopram are acceptable for use; sertraline and mirtazapine are used with caution and paroxetine, fluoxetine, venlafaxine, amitriptyline, imipramine and nortriptyline should be avoided or monitored carefully when used.

The therapeutic regiments associated with patient genotype No. 8 (HTR2A TT/TC; 5HTTR l/l; 1A2, 3A4 and 2C19 are extensive; and 2D6 intermediate) are acceptable for use with citalopram, fluvoxamine, bupropion, escitalopram, sertraline and mirtazapine; paroxetine, fluoxetine, venlafaxine, amitriptyline, imipramine and nortriptyline should be avoided or monitored carefully when used.

The therapeutic drug regimens associated with patient genotype number 9 (HTR2 ATT/TC; 5HTTR l/s; and 1A2, 3A4, 2C19 and 2D6 are extensive) indicate that citalopram, fluvoxamine, bupropion, escitalopram, sertraline, mirtazapine, paroxetine, fluoxetine, venlafaxine, amitriptyline, imipramine and nortriptyline are acceptable for use.

The therapeutic drug history associated with patient genotype 10 (HTR2A TT/TC; 5HTTR l/s; 1A2, 3A4 and 2C19 are extensive; and 2D6 is weak) indicates that citalopram, fluvoxamine, bupropion and escitalopram are acceptable for use, that sertraline and mirtazapine should be used with caution, and that paroxetine, fluoxetine, venlafaxine, amitriptyline, imipramine and nortriptyline should be avoided or monitored carefully when used.

Example 5

The genotypes of the patient CYP2D6, CYP2C19, CYP1a2, and HTR2A genes were determined using the methods described above. Based on the CYP2D6 genotype, patients were classified as either weak, intermediate, extensive or ultrafast metabolic. For the CYP2C19 genotype, patients were classified as either extensive, intermediate or weak metabolic. For genotype 1a2, patients were classified as weak, intermediate, extensive or ultrafast metabolic. For the 5HTTR genotype, patients are classified as having short/short (s/s) form of the gene, short/long (s/l) form of the gene, or long/long (l/l) form of the gene. For the HR2A genotype, patients were classified as having a C/C allele, a T/T allele or a T/C allele.

The following 5 rules were used to sort genotypes as treatment recommendations. The first rule relates to the CYP2D6 genotype. For the weakly and ultrafast metabolic types, the CYP2D6 substrate is placed in a "carefully monitored and carefully used" list. For the intermediate metabolic forms, specialized CYP2D6 substrates were placed in a "cautious use to avoid or closely monitor" list. For the intermediate metabolic forms, substrates that are partially metabolized by CYP2D6 are placed in a "cautious" list. For a wide range of metabolic forms, CYP2D6 substrates are placed in a "use acceptable" list.

After the first rule is implemented, a second rule involving the 2C19 genotype may be implemented. For the weakly metabolic forms based on their CYP2C19 genotype, specialized CYP2C19 substrates were placed in a "avoid or closely monitor cautious" list. For broad or intermediate metabolic types based on their CYP2C19 genotype, CYP2C19 substrates were placed in a "use acceptable" list. Substrates metabolized primarily by both CYP2D6 and CYP2C19 may be placed in a "cautious" list of avoiding or closely monitoring if CYP2C19 is weak and CYP2D6 is moderate or weak, or may be placed in a "cautious" list if CYP2C19 is weak and CYP2D6 is extensive.

The first rule and the second rule are followed by a third rule relating to genotype 1a 2. For weak and ultrafast metabolic types based on their CYP1a2 genotypes, specialized CYP1a2 substrates can be placed in a "carefully used to avoid or closely monitor" list (including fluvoxamine as the primary CYP1a2 substrate). For weak metabolic types based on the CYP1a2 genotype, substrates with the CYP1a2 pathway as one of their major pathways can be placed in a "cautious" list. For broad or intermediate metabolic types based on their CYP1a2 genotype, CYP1a2 substrates may be placed in a "use acceptable" list. In other embodiments, intermediate metabolomics can be placed in a "cautious" list.

After the first three rules were implemented, a fourth rule involving the 5HTTR genotype was implemented. For homozygous short genotypes (s/s) of the 5-hydroxytryptamine transporter gene, SSRIs classified as "acceptable for use" can be transferred to the "caution use" list. For homozygous short genotypes (s/s) of the 5-hydroxytryptamine transporter gene, SSRIs classified as "cautious" can be transferred to a "avoid or closely monitor cautious" list. Antidepressants other than the SSRI class are not based on the 5HTTR genotype transfer list.

After the first four rules are implemented, a fifth rule involving the HR2A genotype is implemented. For the "mutant form" of the 5-hydroxytryptamine receptor 2A gene, if paroxetine is in the "use acceptable" category, it can be transferred to the "caution use" category. If paroxetine is in the "cautious" list, it may be transferred to "avoid or closely monitor very cautious use".

Based on the patient's genotype and associated therapeutic history, the appropriate therapeutic can be selected for the patient.

Example 6

This example describes an electronic illustration of managing patients participating in a pharmaceutical genomics study. Thirty-eight (38) patients were treated with venlafaxine and genotyped for the 2D6 gene. Participants who are slow (also called "weak") metabolic types, defined by having two inactive 2D6 alleles or one inactive and one reduced 2D6 allele, cannot be maintained at doses above 75mg venlafaxine. In contrast, 26 of the remaining 33 subjects with at least one fully functional 2D6 allele (79%) tolerated a venlafaxine dose of 150mg or higher (p <.002).

Clinical guidelines recommend a typical starting dose of venlafaxine to be 75 mg/day. However, the initial dose of 37.5 mg/day is often prescribed for elderly patients. For patients who do not respond to the initial dose of 75 mg/day, it is recommended to increase the daily dose by an increment of 75mg up to a maximum dose of 225 mg/day. Patients with an incompletely active 2D6 gene are expected to have higher serum levels of venlafaxine and N-desmethylvenlafaxine (i.e., inactive metabolites) for any given dose of venlafaxine than patients with one or two functional copies of the 2D6 gene. Thus, a compromised (complexed) metabolic profile is less able to tolerate higher doses of the 2D6 substrate therapeutic. Some patients with limited metabolic capability of 2D6 may be expected to achieve therapeutic clinical response at relatively low doses.

Method

Samples of 199 patients with documented history of atypical response psychotherapeutic drugs were genotyped for the 2D6 gene as part of the clinical study protocol. Atypical responses are defined as the development of adverse side effects or non-response to therapeutic drugs. 14 variant alleles were examined using the TM Bioscience platform. The retest-retest reliability of the 2D6 genotype determination was greater than 99% accuracy. Review of electronic medical records revealed that 38 of the present protocol participants had previously been treated with venlafaxine by clinicians who did not know their 2D6 genotype prior to or during their course of treatment. Table 10 provides the 2D6 alleles and corresponding activity levels.

Watch 10

Alleles	Level of activity
		＊1	Is normal
＊2A	Is raised
		＊2BD	Reduce
＊3	Is free of
		＊4	Is free of
＊5	Is free of
		＊6	Is free of
＊7	Is free of
		＊8	Is free of
＊9	Reduce
		＊10	Reduce
＊11	Is free of
		＊12	Reduce
＊17	Reduce
		＊41	Reduce

5 "slow metabolism" were identified in this group of 38 patients. Each of these patients had a completely inactive allele (e.g.. x 3 or. x 4). In addition, each patient had a second allele with reduced activity (e.g.. times.9,. times.10,. times.17 or. times.41). All other 33 patients treated with venlafaxine had at least one fully active allele.

None of the 5 patients with slow 2D6 metabolism could maintain a dose of venlafaxine higher than 75 mg. In contrast, 26 of the remaining 33 subjects (79%) remained at a dose level of 150mg or 225mg venlafaxine. Using Fisher's exact detection, the difference between dose levels was significant (p <. 002). The following shows a clinical description of 5 subjects lacking a functional 2D6 gene.

Case 1: a44 year old caucasian woman diagnosed with dysthymic disorder was prescribed 75mg venlafaxine. She experienced dry mouth and increased appetite, but reported reduced depression symptoms. Her dose was increased to 112.5mg, at which time her depression was exacerbated, complaining of fatigue. Her dose was then reduced to 75mg, and she responded well to this dose for 4 months. She had the 32D6 allele and the 92D6 allele.

Case 2: a caucasian adolescent age 16 years old diagnosed with attention deficit hyperactivity disorder at age 6. She then became depressed at age 13, developed psychotic symptoms at age 15 and was diagnosed with bipolar disorder at age 16. She was prescribed 75mg of venlafaxine for one month, after which treatment was discontinued because of excessive sleepiness and no improvement in mind. She had the 32D6 allele and the 92D6 allele.

Case 3: a54 year old caucasian woman diagnosed with major depressive disorder was initially prescribed 37.5mg of venlafaxine. After 4 days the treatment was discontinued due to intolerable nausea, insomnia and reduced appetite. She had 42D6 allele and 172D6 allele.

Case 4: a46 year old caucasian male diagnosed with generalized anxiety disorder. He was prescribed 37.5mg of venlafaxine for one day, four days thereafter becoming more anxious. When complaining of increased anxiety, he also developed physiological symptoms including palpitations. After discontinuation of venlafaxine, his anxiety symptoms improved. He had 42D6 allele and 412D6 allele.

Case 5: teenagers, 15 years old asian americans, were diagnosed with major depressive disorder documented as having a history of psychiatric illness with three overdosing attempts for suicide and five hospitalizations. She was prescribed 75mg of venlafaxine despite fear of reporting some mood swings. Venlafaxine was discontinued after 5 days due to intolerable side effects. She previously experienced side effects when treated with low doses of other 2D6 substrate treatment drugs (e.g., paroxetine and fluoxetine). She had 42D6 allele and 102D6 allele.

Analysis of clinical samples from psychotic patients indicated that individuals lacking a fully functional copy of the 2D6 gene did not tolerate a maintenance dose of venlafaxine higher than 75 mg. In one case, a subject can take 112.5mg shortly before the treating clinician reduces the dose back to 75mg (case 5). This is the only patient who has an inactive and partially inactive 2D6 allele and who tolerates any dose of venlafaxine. In contrast, 79% of patients with at least one fully functional copy of the 2D6 gene tolerate doses of 150mg or more of venlafaxine.

Two patterns were confirmed when reviewing 5 patients identified as "slow metabolism". Venlafaxine was not tolerated in 4 patients due to a range of side effects including nausea, insomnia, anorexia, palpitations and lethargy. Subject 1, named "slow metabolizing", had a therapeutic response at 75mg, but she did not tolerate a dose up to 112.5 mg.

One clinical implication of these analyses is: for patients lacking a functional copy of the 2D6 allele, venlafaxine should start at a dose of 37.5mg, with cautious increases in the dose. Another strategy is to select an alternative antidepressant whose primary metabolic pathway is not 2D6, such as fluvoxamine, or an antidepressant with an alternative metabolic pathway, such as escitalopram or sertraline.

The strength of this clinical report is that none of the treating psychiatrists who treated these patients recognized their 2D6 genotype. Thus, dosing decisions are based on the clinical response of the patient rather than predicting a phenotype based on genotypic information.

One drawback of this report is that clinical data is retrieved from electronic medical records. Although venlafaxine doses were always accurately recorded, no efficacy descriptions and patient response to treatment were reported in a standard manner. Inclusion criteria involved in this study confirmed an atypical response to antidepressant therapeutic drugs, defined by the development of adverse side effects or non-response to the therapeutic drugs. Thus, the decision to maintain, increase or decrease the dosage of a therapeutic agent is based on the patient's tolerance and response to the therapeutic agent, regardless of diagnosis and concomitant disease.

In summary, patients who do not have at least one fully functional allele of the 2D6 gene do not maintain venlafaxine doses greater than 75 mg. The correlation of this compromised genotype with the clinical history of intolerance to higher doses of venlafaxine indicates that genetic variation in cytochrome P4502D6 may cause specific changes in patients observed over time in response to antidepressant drug therapy.

Other embodiments

It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Sequence listing

<110> D.A. Marazake (Mrazek, David A.) D.J. Okaien (O' Kane, Dennis J.) J.L. Blackiii (Black III, John L.)

<120> method for selecting therapeutic drugs

<130>07039/718001

<140>11/498,976

<141>2006-08-02

<150>10/545,931

<151>2005-08-18

<150>60/448,547

<151>2003-02-20

<160>180

<170>FastSEQ for Windows Version 4.0

<210>1

<211>22

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>1

ccagaaggct ttgcaggctt ca 22

<210>2

<211>22

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>2

actgagccct gggaggtagg ta 22

<210>3

<211>31

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>3

ggcctaccct gggtaagggc ctggagcagg a 31

<210>4

<211>17

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>4

cctggtcgaa gcagtat 17

<210>5

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>5

gcacgctacc 10

<210>6

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>6

gcacgctact 10

<210>7

<211>20

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>7

caccaggccc cctgccactg 20

<210>8

<211>20

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>8

cctgctcact cctggtagcc 20

<210>9

<211>19

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>9

ctgtttcatg tccacgacc 19

<210>10

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>10

tgcccatcac 10

<210>11

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>11

tgcccatcat 10

<210>12

<211>24

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>12

ccagatcctg ggtttcgggc cgcg 24

<210>13

<211>16

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>13

cagaggcgct tctccg 16

<210>14

<211>20

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>14

ctcggtctct cgctccgcac 20

<210>15

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>15

cttctccgtg 10

<210>16

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>16

cttctccgtc 10

<210>17

<211>26

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>17

tccaccttgc gcaacttggg cctggg 26

<210>18

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>18

ccacccccag 10

<210>19

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>19

ccacccccaa 10

<210>20

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>20

gacgcccctt tcgccccaac g 21

<210>21

<211>20

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>21

tgagacttgt ccaggtgaac 20

<210>22

<211>20

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>22

cccagatggg ctcacgctgc 20

<210>23

<211>10

<212>DNA

<213> Artificial sequence

<400>23

actgagcaca 10

<210>24

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>24

actgagcacg 10

<210>25

<211>25

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>25

ggatgacctg ggacccagcc cagcc 25

<210>26

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>26

gagaacctgc 10

<210>27

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>27

gagaacctgt 10

<210>28

<211>27

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>28

gcatagtggt ggctgacctg ttctctg 27

<210>29

<211>20

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>29

cccccgctga ctcccctctg 20

<210>30

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>30

gacccccgag cctcaccttc c 21

<210>31

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>31

agctgccacc 10

<210>32

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>32

agctgccact 10

<210>33

<211>15

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>33

gcggaggccc caggt 15

<210>34

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>34

accaagaggg aagaagcaca g 21

<210>35

<211>20

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>35

ccaatgacgg acaggagaaa 20

<210>36

<211>11

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>36

gagctggtga g 11

<210>37

<211>11

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>37

gagctggtga a 11

<210>38

<211>20

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>38

ctgcactccg ttctgctcct 20

<210>39

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>39

gatgggggtc ctggtatgtc t 21

<210>40

<211>19

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>40

ctggaggtca cggctcaag 19

<210>41

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>41

gtgtggcatg gaccttgtct g 21

<210>42

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>42

gcaatgcgcc gtatttgttt c 21

<210>43

<211>15

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>43

aaagcttatt acaga 15

<210>44

<211>13

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>44

agcttattac agc 13

<210>45

<211>23

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>45

ggatgagatg gcatatgtcc tgc 23

<210>46

<211>20

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>46

cctgtttcct gtcgctgctc 20

<210>47

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>47

ccaatacctg tccacgctga t 21

<210>48

<211>11

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>48

ggtgtcggaa c 11

<210>49

<211>11

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>49

ggtgtcggaa a 11

<210>50

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>50

ctgataacgg cagcacagac c 21

<210>51

<211>17

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>51

gggctggtgg tggagag 17

<210>52

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>52

tctgacacag ccaaggagat g 21

<210>53

<211>9

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>53

ccaggagcg 9

<210>54

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>54

cccaggagcc 10

<210>55

<211>19

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>55

tggacaggat gagcagcga 19

<210>56

<211>17

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>56

gggctggtgg tggagag 17

<210>57

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>57

tctgacacag ccaaggagat g 21

<210>58

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>58

ccaggagcgt 10

<210>59

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>59

ccaggagcgg 10

<210>60

<211>20

<212>DNA

<213> Artificial sequence

<400>60

ggacaggatg agcagcgaca 20

<210>61

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>61

tctccacagc actcccgaca g 21

<210>62

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>62

ccgagaacaa tggcgagcat c 21

<210>63

<211>9

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>63

ggtccgggt 9

<210>64

<211>9

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>64

ggtccgggc 9

<210>65

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>65

tttgccattg ggcatggtct g 21

<210>66

<211>18

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>66

caccagccca cccgagag 18

<210>67

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>67

accgagaaca atggcgagca t 21

<210>68

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>68

ccatggtggg 10

<210>69

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>69

ccatggtggc 10

<210>70

<211>17

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>70

acgggtcggg gagagtc 17

<210>71

<211>18

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>71

ccggtacagc cccatccc 18

<210>72

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>72

gccgactcac cgagaacaat g 21

<210>73

<211>9

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>73

gtgggacgg 9

<210>74

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>74

ggtgggacga 10

<210>75

<211>19

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>75

gtcggggaga gtcagctgg 19

<210>76

<211>22

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>76

ggggacagga gggagggttg ag 22

<210>77

<211>25

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>77

gaagaggagt gggcaggaga tggtg 25

<210>78

<211>12

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>78

ggagatcatg ac 12

<210>79

<211>13

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>79

tggagatcat gat 13

<210>80

<211>16

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>80

ggtgacccgg cgcttg 16

<210>81

<211>20

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>81

agcaaccaag ccccaaagag 20

<210>82

<211>19

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>82

gagcgcgcag taggagagg 19

<210>83

<211>11

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>83

ttcaggtggc t 11

<210>84

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>84

tcaggtggcc 10

<210>85

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>85

actcagctgg ctcagagatg c 21

<210>86

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>86

cgctacaacc ggcagggtgg g 21

<210>87

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>87

gcagcatgag cgggcagggt a 21

<210>88

<211>11

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>88

ggacgagtag a 11

<210>89

<211>11

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>89

ggacgagtag c 11

<210>90

<211>19

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>90

ccacgtagtc gcggtcctc 19

<210>91

<211>19

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>91

cgccttcccc cacgccacc 19

<210>92

<211>20

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>92

gttggagccg caggggtccg 20

<210>93

<211>8

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>93

ggcgcggg 8

<210>94

<211>8

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>94

ggcgcggc 8

<210>95

<211>15

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>95

gggcgcacag tcggg 15

<210>96

<211>20

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>96

tgcggttttc catcaagaag 20

<210>97

<211>18

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>97

ggagttggcc cagcagaa 18

<210>98

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>98

ccgacagggt 10

<210>99

<211>9

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>99

cgacagggg 9

<210>100

<211>27

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>100

cttgagaacc ttggtctcct tcttgat 27

<210>101

<211>18

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>101

caacgccgac ttctggaa 18

<210>102

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>102

acatgcgatc gaaaggactc t 21

<210>103

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>103

cggggggaac 10

<210>104

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>104

cggggggaat 10

<210>105

<211>19

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>105

ggcgttgggc atcatgtgg 19

<210>106

<211>18

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>106

gcagtccagc ccgaaatg 18

<210>107

<211>16

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>107

gcacgatggc tgcgga 16

<210>108

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>108

ctgcggacac 10

<210>109

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>109

ctgcggacat 10

<210>110

<211>15

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>110

cagcacgttg cccag 15

<210>111

<211>13

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>111

caacgtgctg gtg 13

<210>112

<211>15

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>112

catcttgcgc ttgta 15

<210>113

<211>8

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>113

ggctgcgc 8

<210>114

<211>9

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>114

tggctgcgg 9

<210>115

<211>17

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>115

acaccagcac gttgccc 17

<210>116

<211>16

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>116

cgccaagatg accaac 16

<210>117

<211>17

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>117

cggaatgaag gagatga 17

<210>118

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>118

gccacgaaga 10

<210>119

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>119

gccacgaagc 10

<210>120

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>120

ggtctgacac agccagagac a 21

<210>121

<211>20

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>121

gcagctgcct acatccacat 20

<210>122

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>122

cattcggggt gaaaggtgtt a 21

<210>123

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>123

gtccagctcc 10

<210>124

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>124

gtccagctct 10

<210>125

<211>23

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>125

cagacagact ctgcaacagg gtc 23

<210>126

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>126

ggagagtcgg ggagcaggaa c 21

<210>127

<211>17

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>127

gaggcggggg cacaagg 17

<210>128

<211>11

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>128

gcacagagca c 11

<210>129

<211>11

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>129

gcacagagca t 11

<210>130

<211>18

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>130

ccccagcctt gctctcca 18

<210>131

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>131

ctacatcccg ctgctgctca t 21

<210>132

<211>19

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>132

cctgctcccc gactctcca 19

<210>133

<211>9

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>133

gcgcagctc 9

<210>134

<211>9

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>134

gcgcagcta 9

<210>135

<211>24

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>135

ggaatatgcg cccatagaga acca 24

<210>136

<211>25

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>136

gctcacttgg cttattggct tcctc 25

<210>137

<211>20

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>137

catgagcagc agcgggatgt 20

<210>138

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>138

catgcgtcgg 10

<210>139

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>139

catgcgtcga 10

<210>140

<211>18

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>140

ggtccgagcg gtcttccg 18

<210>141

<211>24

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>141

agggcaacaa caccacatca ccac 24

<210>142

<211>22

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>142

gcagcaccaa caccgacacc at 22

<210>143

<211>11

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>143

acgtcggaga t 11

<210>144

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>144

cgtcggagac 10

<210>145

<211>19

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>145

accagtagtg ttgccgccg 19

<210>146

<211>20

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>146

tcagggcaac aacaccacat 20

<210>147

<211>18

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>147

gcaccaacac cgacacca 18

<210>148

<211>9

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>148

gttgccgcc 9

<210>149

<211>9

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>149

gttgccgct 9

<210>150

<211>19

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>150

ggtctcaaag ggagccggt 19

<210>151

<211>20

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>151

ttttcttccc tcccccttcc 20

<210>152

<211>17

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>152

ccaccacgca cgcattg 17

<210>153

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>153

aagggagccg 10

<210>154

<211>11

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>154

aaagggagcc a 11

<210>155

<211>22

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>155

gtggtgatgt ggtgttgttg cc 22

<210>156

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>156

gctcaactac gaactcccta a 21

<210>157

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>157

tcactacggc tgtcagtaaa g 21

<210>158

<211>12

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>158

ttagcttctc ca 12

<210>159

<211>12

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>159

ttagcttctc cg 12

<210>160

<211>29

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>160

gagttaaagt cattactgta gagcctggt 29

<210>161

<211>16

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>161

gctctgctcg ctcaac 16

<210>162

<211>21

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>162

gattttagtg gctttccttt c 21

<210>163

<211>9

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>163

ccgagtgcg 9

<210>164

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>164

gccgagtgca 10

<210>165

<211>18

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>165

agtgcccctc atggaggc 18

<210>166

<211>25

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>166

ctcaatttta aactttggtt gctta 25

<210>167

<211>23

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>167

ttttgaatag gaaacaccca taa 23

<210>168

<211>11

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>168

gcattcctca g 11

<210>169

<211>11

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>169

gcattcctca c 11

<210>170

<211>28

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>170

gttcaccatg attgcttcag tcttaagc 28

<210>171

<211>30

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>171

atgatgacaa tgatgctgat gatgataatg 30

<210>172

<211>25

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>172

tccaccatcg gaggtattga aaatg 25

<210>173

<211>15

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>173

tcacagaaat atcac 15

<210>174

<211>14

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>174

cacagaaata tcag 14

<210>175

<211>26

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>175

attgccaaac caataggcca attagg 26

<210>176

<211>20

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>176

ggcttcagtt tcccgatctg 20

<210>177

<211>20

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>177

taggcaaggg caggcactac 20

<210>178

<211>10

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>178

aaggagctgg 10

<210>179

<211>11

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>179

gaaggagctg a 11

<210>180

<211>16

<212>DNA

<213> Artificial sequence

<220>

<223> oligonucleotide

<400>180

gtacatgggg gccccc 16

Claims (22)

1. A method of selecting a psychotherapeutic drug for a patient, the method comprising:

(a) providing the genotype of a panel of genes of the patient, wherein the panel of genes comprises a CYP2D6 gene, a CYP2C19 gene, a CYP1A2 gene, a 5-hydroxytryptamine transporter gene, and a 5-hydroxytryptamine receptor 2A gene; and are

(b) Selecting the psychotherapeutic drug based on the genotype.

2. The method of claim 1, wherein providing the patient's genotype comprises determining whether the patient comprises a CYP1A 2x 1A or 1A 2x 3 allele, CYP2C19 x 1A, 2C19 x 1B, or 2C19 x 2A allele and a CYP2D6 x 1A, 2D6 x 2, 2D6 x 2N, 2D6 x 3, 2D6 x 4, 2D6 x 5, 2D6 x 6, 2D6 x 7, 2D6 x 8, 2D6 x 10, 2D6 x 12, or 2D6 x 17 allele.

3. The method of claim 1, wherein providing the patient's genotype further comprises determining whether the patient comprises the 2D6 x 41 allele.

4. The method of claim 1, wherein the set of genes comprises 4 cytochrome P450 genes.

5. The method of claim 4, wherein the 4 cytochrome P450 genes are CYP2D6, 2C19, 3A4, and 1A 2.

6. The method of claim 5, wherein providing the patient's genotype comprises determining whether the patient comprises CYP2D 6X 2, 2D 6X 3, 2D 6X 4, 2D 6X 10 or 2D 6X 17 allele, CYP2C 19A 2C 19X 2B, 2C19 3, 2C 19X 4, 2C 19X 5A, 2C 19X 5B, 2C 19X 6, 2C 19X 3 or 2C 19X 8 allele, CYP3A 4X 1B, 3A 4X 2, 3A 4X 5, 3A 4X 6, 3A 4X 12, 3A 4X 13, 3A 4X 2, 3A 5943A 863A, and CYP 8427X 3A 8617A.

7. The method of claim 1, wherein the set of genes comprises 5 cytochrome P450 genes.

8. The method of claim 7, wherein the 5 cytochrome P450 genes are CYP1A1, 1A2, 2D6, 2C19, and 3A 4.

9. The method of claim 8, wherein providing the patient's genotype comprises determining whether the patient comprises a CYP1A 1A1, 1A 1A2, 1A 1A 3 or 1A 1A 4 allele, a CYP1A 2A or 1A2 A3 allele, a CYP2C19 a, 2C 19B or 2C19 a allele, a CYP2D 6a 1, 2D 6a 2D6 2N, 2D 6A3, 2D6 x 4, 2D6 A5, 2D6 A6, 2D6 A7, 2D6 A8, 2D 6310, 2D 6a 68612, 2D 5812A 862A 863 or a 82863 allele.

10. The method of claim 1, wherein the set of genes further comprises a dopamine transporter gene.

11. The method of claim 1, wherein the set of genes further comprises a plurality of dopamine receptor genes.

12. The method of claim 11, wherein the dopamine receptor gene encodes dopamine receptors D1, D2, D3, D4, D5, and D6.

13. The method of claim 1, wherein the set of genes further comprises a 5-hydroxytryptamine receptor gene encoding 5-hydroxytryptamine receptor 1A, 1B, 1D, or 2C.

14. The method of claim 1, wherein the set of genes further comprises a catechol-o-methyltransferase gene.

15. The method of claim 1, wherein the set of genes comprises at least 10 cytochrome P450 genes.

16. The method of claim 15, wherein the at least 10 cytochrome P450 genes are CYP1a1, 1a2, 1B1, 2A6, 2B6, 2C8, 2C9, 2C18, 2C19, 2D6, 2E1, 3a4, and 3a 5.

17. The method of claim 16, wherein providing the patient's genotype comprises determining whether the patient comprises a CYP1A 1x 1A, 1A 1x 2, 1A 1x 3, or 1A 1x 4 allele; 1A 2x 1A or 1A 2x 3 allele; CYP1B 1x 1, 1B 1x 2, 1B 1x 3, 1B 1x 4, 1B 1x 11, 1B 1x 14, 1B 1x 18, 1B 1x 19, 1B 1x 20, or 1B 1x 25 alleles; CYP2a6 x 1A, 2a6 x 1B, 2a6 x 2 or 2a6 x 5 allele; CYP2B6 x 1, 2B6 x 2, 2B6 x 3, 2B6 x 4, 2B6 x 5, 2B6 x 6, or 2B6 x 7 alleles; CYP2C8 x 1A, 2C8 x 1B, 2C8 x 1C, 2C8 x 2, 2C8 x 3, or 2C8 x 4 alleles; CYP2C9 x 1, 2C9 x 2, 2C9 x 3 or 2C9 x 5 allele; CYP2C18 m1 or 2C18 m2 alleles; CYP2C19 x 1A, 2C19 x 1B or 2C19 x 2A alleles; CYP2D 6a, 2D 6a 2, 2D 6a 2N, 2D 6a3, 2D 6a 4, 2D 6a 5, 2D 6a 6, 2D 6a 7, 2D 6a 8, 2D 6a 10, 2D 6a 12, 2D 6a 17 or 2D 6a 35 allele; CYP2E 1x 1A, 2E 1x 1C, 2E 1x 1D, 2E 1x 2, 2E 1x 4, 2E 1x 5 or 2E 1x 7 alleles; CYP3a4 x 1A or 3a4 x 1B allele; and CYP3a5 x 1A, 3a5 x 3, 3a5 x 5 or 3a5 x 6 alleles.

18. The method of claim 1, wherein the set of genes further comprises a tryptophan hydroxylase gene.

19. The method of claim 1, wherein the psychotropic drug is an antidepressant.

20. The method of claim 1, wherein the psychotropic medication is an antipsychotic, an anxiolytic-sedative, or a mood stabilizer.

21. The method of claim 1, wherein selecting the psychotropic medication comprises:

(a) correlating the genotype of said cytochrome P450 genes with the ability of each cytochrome P450 enzyme encoded by each said cytochrome P450 gene to metabolize said psychotherapeutic drug; and are

(b) Correlating the genotype of the 5-hydroxytryptamine transporter gene and the 5-hydroxytryptamine receptor 2A gene with the patient's ability to respond to the psychotherapeutic drug.

22. An article comprising a substrate comprising a plurality of discrete regions, wherein each of said regions comprises a different population of nucleic acid molecules, wherein said different populations of nucleic acid molecules independently comprise nucleic acid molecules for detecting: CYP1A 2x 1A and 1A 2x 3 alleles; CYP2C19 x 1A, 2C19 x 1B, and 2C19 x 2A alleles; CYP2D 6a, 2D 62, 2D 6N, 2D 63, 2D 64, 2D 65, 2D 66, 2D 67, 2D 68, 2D6 10, 2D6 12, 2D6 17 and 2D 635 alleles; the 5-hydroxytryptamine receptor 2A C/T, T/T, C/C allele; and 5HTT promoter repeat and exon 2 variable repeat alleles.