Kit and method for detecting tamoxifen personalized medicine genetic polymorphism by use of pyrosequencing technique

A pyrosequencing method and gene polymorphism technology, applied in the field of molecular biology, can solve problems such as chemotherapy effect and survival rate reduction, and achieve the effect of simple operation and small sample size

Inactive Publication Date: 2012-08-22
CENT SOUTH UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, if the product cannot be formed, the efficacy of chemotherapy and the survival rate will be greatly reduced

Method used

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  • Kit and method for detecting tamoxifen personalized medicine genetic polymorphism by use of pyrosequencing technique
  • Kit and method for detecting tamoxifen personalized medicine genetic polymorphism by use of pyrosequencing technique
  • Kit and method for detecting tamoxifen personalized medicine genetic polymorphism by use of pyrosequencing technique

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Embodiment 1

[0033] CYP2D6*10 (rs1065852) upstream primer: 5′-GTA GTG AGG CAG GTA TGG G-3′ (SEQ ID NO.3);

[0034] SULT1A1*2 (rs9282861) upstream primer: 5′-GGA GAT TCA AAA GAT CCTGGA-3′ (SEQ ID NO.4);

[0035] CYP2D6*10 (rs1065852) downstream primer: 5'-CAG GAC CTC CTC CCT CAC CTGGTC-3' (SEQ ID NO.5);

[0036] SULT1A1*2 (rs9282861) downstream primer: 5′-TTC ATC TCC TTG AAC GAC G-3′ (SEQ ID NO.6);

[0037] CYP2D6*10 (rs1065852) sequencing primer: 5'-GCA GGG GGC CTG GTG-3' (SEQ ID NO.7);

[0038] SULT1A1*2 (rs9282861) sequencing primer: 5′-TGG AGT TTG TGG GGC-3′ (SEQID NO.8);

[0039] 1. DNA extraction

[0040] 1.1 The preparation and inspection of reagent materials before the experiment are as follows:

[0041] (1) Check the shelf life of the kit and ensure that ethanol has been added to Wash Buffer 1 and 2, and tick the corresponding mark on the bottle; (2) Isopropanol (if not available, it can be replaced by absolute ethanol) and 75% ethanol ; (3) 1.5mL Eppendorf tubes and various p...

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Abstract

The invention discloses a kit and method for detecting the tamoxifen personalized medicine genetic polymorphism by use of the pyrosequencing technique. The genetic polymorphism specifically refers to the single nucleotide polymorphism of CYP2D6*10(rs1065852) and (SULT1A1*2) (rs9282861). The kit comprises the primer shown by SEQ ID NO.3-8. The kit disclosed by the invention can realize accurate, quick and high-flux detection on CYP2D6*10(rs1065852) and (SULT1A1*2) (rs9282861) so as to realize safe, reasonable and effective personalized administration of the tamoxifen medicine.

Description

technical field [0001] The invention belongs to the field of molecular biology, and specifically relates to a kit and a method for detecting gene polymorphism of tamoxifen individualized drug by pyrosequencing method. Background technique [0002] Tamoxifen is a non-steroidal anti-estrogen drug, which is currently widely used in the prevention and treatment of estrogen receptor-positive breast cancer patients. The metabolism of tamoxifen in the body involves a variety of drug-metabolizing enzymes, and about 10% of tamoxifen is metabolized by CYP2D6 into the active metabolite 4-hydroxy-N-desmethyl tamoxifen (endoxifen). In the Chinese population, the most common CYP2D6 loss-of-function allele mutation is CYP2D6*10 (about 53%). In a study of 200 breast cancer patients receiving TAM as an adjuvant therapy, CYP2D6 slow metabolizers had a lower disease-free survival rate, which is consistent with the slowed production of the active metabolite endoxifen, which leads to a decline ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 周宏灏
Owner CENT SOUTH UNIV
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