Kit for detecting glutathione S-transferase pi 1 (GSTP1) gene polymorphism by using pyrosequencing method and method

A technology of pyrosequencing and gene polymorphism, applied in the field of molecular biology, can solve the problems of reducing enzyme activity, poor curative effect, and reducing the activation efficiency of anthracyclines, and achieves the effect of small sample volume and easy operation.

Inactive Publication Date: 2012-11-14
周宏灏
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  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

The occurrence of A>G at its rs1695 site will cause Ile105Val, thereby reducing the activity of the enzyme, resulting in a decrease in the activation efficiency of anthracyclines
Carrying A / A (70.5%) genotype has poor curative effect

Method used

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  • Kit for detecting glutathione S-transferase pi 1 (GSTP1) gene polymorphism by using pyrosequencing method and method
  • Kit for detecting glutathione S-transferase pi 1 (GSTP1) gene polymorphism by using pyrosequencing method and method
  • Kit for detecting glutathione S-transferase pi 1 (GSTP1) gene polymorphism by using pyrosequencing method and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] GSTP1-pyroF (upstream primer): 5'- CTG GTG GAC ATG GTG AAT -3' (SEQ ID NO.2);

[0031] GSTP1-pyroR (downstream primer): 5'- CCC AGT GCC CAA CCC TGG -3' (SEQ ID NO.3);

[0032] Sequencing primer: 5'-CCT CCG CTG CAA ATA -3' (SEQ ID NO.4);

[0033] 1. DNA extraction

[0034] 1.1 The preparation and inspection of reagent materials before the experiment are as follows:

[0035] (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 marks on the bottle; (2) Isopropanol (if not available, absolute ethanol can be used instead) and 75% ethanol ; (3) 1.5mL Eppendorf tubes and various pipette tips within the validity period of autoclaving.

[0036] 1.2 Take out the EDTA anticoagulant tube containing whole blood from the refrigerator at 4°C, and mix it upside down several times;

[0037] 1.3 Make a mark on the 1.5mL Eppendorf tube corresponding to the unique identification of the specimen;

[0038] 1.4 Pipe...

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Abstract

The invention discloses a kit for detecting GSTP1 gene polymorphism by using a pyrosequencing method and the method. The GSTP1 gene polymorphism refers to the rs1695 (A>G) single nucleotide polymorphism in particular. The kit comprises a primer represented by SEQ ID NO.2-4. The kit can achieve the accurate, rapid and high-throughput detect the GSTP1 gene polymorphism, so that safe, reasonable and effective individuation dosing can be achieved to substrates of platinum chemotherapy drugs, anthracycline antibiotics and the like.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a kit and a method for detecting GSTP1 gene polymorphism by pyrosequencing. Background technique [0002] Human glutathione S-transferase P1 (GSTP1) is the most important phase II detoxification enzyme in the body. It mainly catalyzes the combination of GSH and the active metabolites of various exogenous chemicals in the body. It is involved in the detoxification and metabolism of many cancer chemotherapy drugs. makes an important impact. [0003] Platinum anticancer drugs are the first-line chemotherapeutic drugs used in the chemotherapy of various tumors. The common ones are cisplatin, carboplatin, oxaliplatin, nedaplatin, and lobaplatin. The GSTP1 rs1695 (Ile105Val) mutation leads to a significant decrease in enzyme activity, reduces the metabolic clearance rate of platinum-based chemotherapy drugs, and prolongs the effect of platinum-based chemotherapy drugs on ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 周宏灏
Owner 周宏灏
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