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Clozapine individualized drug gene detection kit

A gene detection and kit technology, which is applied in the field of gene detection kits for detection of clozapine individualized drug use, can solve the problem of lack of detection sensitivity, high accuracy, high clozapine individualized drug gene detection kits, and slow metabolism of antipsychotic drugs , high blood concentration and other problems, to achieve the effect of reducing medical expenses, reducing drug side effects, and high sensitivity

Pending Publication Date: 2019-01-15
南通中科医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that CYP2D6 is the main metabolic enzyme of clozapine, and the metabolism of antipsychotic drugs in this genotype is slower and the blood concentration is higher, resulting in more obvious side effects
[0003] The prior art mainly has the following technical problems: lack of a gene detection kit for clozapine individualized drug use aimed at the Chinese population with high detection sensitivity and high accuracy

Method used

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  • Clozapine individualized drug gene detection kit
  • Clozapine individualized drug gene detection kit
  • Clozapine individualized drug gene detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Genetic detection kit for one-person detection of clozapine individualized medicine, including the following components:

[0040] (1) 4 pairs of PCR amplification and sequencing primers for CYP2D6*2(886C>A), CYP2D6*41(985+39G>A), CYP2D6*10(100C>T) and MC4R(57882787C>T), 1 pair of CYP2D6Z large fragment PCR amplification primers; each primer 1OD,

[0041] The nucleotide sequence of each primer is shown in SEQ NO.3-12, see Table 1 for details:

[0042] Table 1

[0043]

[0044]

[0045] (2) PCR amplification reagent: 15 μL of Taq enzyme mixture (dNTP, 10×PCR reaction buffer, MgCl 2 ); (purchased from Novozyme)

[0046](3) PCR product purification reagent: 3.2 μL SAP enzyme mixture (SAP enzyme, ExoI enzyme, deionized water); (purchased from Novizyme)

[0047] (4) Sequencing reagents: 4.4 μL BigDye mix (Bigdye, 5×seq), 5 μL EDTA, 60 μL ethanol solution (100%), 200 μL ethanol solution (70%), 16 μL Hi-Di.

[0048] This kit is stored at -20°C, and repeated freezing ...

Embodiment 2

[0050] The steps of using the one-person detection clozapine individualized drug gene detection kit of Example 1 include:

[0051] (1) extract the genomic DNA of the sample;

[0052] (2) PCR amplification of CYP2D6*2(886C>A), CYP2D6*41(985+39G>A), CYP2D6*10(100C>T) and MC4R(57882787C>T) loci

[0053] For CYP2D6*2 (886C>A), CYP2D6*41 (985+39G>A), CYP2D6*10 (100C>T) and MC4R (57882787C>T) loci, design primers according to Table 1 for PCR amplification.

[0054] Due to the presence of pseudogenes in the CYP2D6 gene, the kit uses nested PCR amplification. First, use the CYP2D6Z large fragment PCR amplification primers for large fragment amplification, and then amplify CYP2D6*2 / *41 / * on the basis of the amplified products. 10 positions, thereby improving the specificity of the reaction;

[0055] Each reaction system has a total volume of 16 μL, including 7.5 μL of Taq enzyme mixture (dNTP, 10×PCR reaction buffer, MgCl 2 ), deionized water 5.5 μL, primer pair (10uM) 1 μL, genomic...

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Abstract

A clozapine individualized drug gene detection kit comprises the following components: CYP2D6*2 (886C) A, CYP2D6*41 (985+39G) A, CYP2D6*10 (100C) T and MC4R (57882787C) T; 4 pair of site amplificationand sequencing prim, 1 pair of CYP2D6Z large fragment PCR amplification primers, PCR amplification reagent, PCR product purification reagent and DNA sequencing reagent; as shown in Table 1 of that prim sequence. The invention has the following technical effects: a gene detection kit for individualized clozapine medication for Chinese people is provided, and the sensitivity is high and the accuracy is high. The kit is used for CYP2D6*2 (886C) A), CYP2D6*41(985+39G)A), CYP2D6*10 (100C) T) and MC4R (57882787C) T). The genetic difference between individuals can be found through the test of this item, and the drug constitution (slow metabolism or normal metabolism) can be defined, the relevant gene information can be interpreted, and the physiological status or drug reaction can be known in time, so that the drug efficacy can be improved, the toxic and side effects of drugs can be reduced, and the medical cost can be reduced.

Description

technical field [0001] The invention belongs to the field of biological technology, in particular to a gene detection kit for detecting clozapine individualized medicine. Background technique [0002] Clozapine belongs to the diazepine class and is the earliest typical antipsychotic drug. It has a powerful sedative-hypnotic effect and also has a muscarinic effect. It has obvious curative effect on controlling hallucinations, delusions, and restless schizophrenia, and also has certain curative effect on schizophrenia with anxiety or negative symptoms. Clozapine is the only drug approved by the FDA for the treatment of refractory schizophrenia, but its idiopathic and dose-dependent severe side effects limit the clinical application of the drug. In practical application, there are great individual differences in the clinical curative effect of clozapine, and some patients have insignificant curative effect, and even serious adverse reactions. However, there is no effective pre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/6848
CPCC12Q1/6848C12Q1/6883C12Q2600/106C12Q2600/156C12Q2549/119C12Q2535/122
Inventor 陆军王斐景丹丹姜昕
Owner 南通中科医学检验实验室有限公司
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