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Kit for detecting mercaptopurine methyltransferase genetype and method thereof

A technology of mercaptopurine methyl and transferase, which is applied in biochemical equipment and methods, measuring devices, microbiological determination/inspection, etc. It can solve the problem of lack, inability to obtain individualized genetic information of patients in time, low efficiency of enzyme digestion, etc. question

Active Publication Date: 2008-07-02
深圳泰乐德医疗有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the reported PCR-RFLP method has the disadvantages of low amplification efficiency, unstable system, low enzyme digestion efficiency, time-consuming and labor-intensive. Therefore, there has been no mature product using PCR-RFLP method to identify genotypes.
Due to the lack of efficient, stable, convenient and inexpensive methods and products for detecting TPMT genotypes, clinicians cannot obtain the corresponding individualized genetic information of patients in a timely manner when administering drugs, and thus cannot predict the response of different patients to purine according to the results of pharmacogenomics research. Efficacy and side effects of similar drugs, it is difficult to carry out individualized treatment

Method used

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  • Kit for detecting mercaptopurine methyltransferase genetype and method thereof
  • Kit for detecting mercaptopurine methyltransferase genetype and method thereof
  • Kit for detecting mercaptopurine methyltransferase genetype and method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0096] The present invention will be further described in detail below by taking the TPMT460 wild-type positive control plasmid as an example.

[0097] 1. Primers and templates

[0098] Gene fragment amplification using the following primers

[0099] TPMT 460 For: 5'-CAGCGTAGTTGGTGTGGAAA-3';

[0100] TPMT 460Back: 5'-GCGATCACCTGGATTAATGG-3'.

[0101] The template uses human genomic DNA (No. AI011611, concentration 15ng / μl)

[0102] 2. Gene fragment amplification

[0103] Configure the PCR amplification mixture according to Table 3

[0104] Table 3 PCR reaction component preparation

[0105]

[0106] The PCR amplification conditions are:

[0107] 94°C for 3min; 94°C for 30sec, 63°C for 45sec, 72°C for 30sec, 35 cycles; 72°C for 7min, a band with a size of 240bp was obtained after PCR amplification, the results are shown in the appendix figure 1

[0108] 3. Purification of PCR products

[0109] 1). PCR products were subjected to 1.5% agarose gel electrophoresis, the ...

Embodiment 2

[0134] The present invention will be further described in detail below by taking the TPMT genotype detection process of leukemia patient genes as an example.

[0135] Leukemia is a malignant disease of the hematopoietic system, commonly known as "blood cancer", and is one of the top ten high-incidence malignant tumors in China. It is characterized by malignant proliferation of a certain type of leukemia cells in the hematopoietic tissue in the bone marrow or other hematopoietic tissues, and infiltrates the body Various organs and tissues lead to the inhibition of normal hematopoietic cells, resulting in various symptoms. The clinical manifestations are characterized by fever, bleeding, anemia, hepatosplenomegaly, and lymphadenopathy. Leukemia is generally divided into acute and chronic according to the natural course of the disease and the degree of immaturity of the cells. According to the cell type, it is divided into granulocytes, lymphocytes, monocytes and other types. The ...

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PUM

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Abstract

The invention relates to a method and a kit for detecting genotype of thiopurine methyltransferase (TPMT), belonging to gene technology field. The method comprises the following steps of: performing polymerase chain reaction to a sample DNA using specific primers of TPMT gene polymorphism sites, digesting and amplifying the product with restriction endonuclease, and determining genotype according to the length polymorphism of the digested fragments, wherein the primers at least include a specific primer for detecting the TPMT G460A polymorphism site and a specific primer for detecting the A719G polymorphism site. The inventive method and the kit for genotype detection can provides guidance for the individualized administration of purine drugs; and has the advantages of good stability and reliability, low price, rapidness, and applicability to clinic application.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a mercaptopurine methyltransferase genotype detection kit and method. More specifically, the present invention relates to a polymorphism analysis kit and a polymorphism of a gene locus encoding thiopurine methyltransferase (TPMT) by polymerase chain reaction-restriction fragment length polymorphism method method. Background technique [0002] Purine drugs, including 6-mercaptopurine (6-MP), 6-mercaptopurine (6-TG) and mercaptopurine (AZA) are commonly used clinical tumor chemotherapy drugs, and are widely used in the treatment of various types of leukemia. In addition to leukemia, purine drugs are also widely used in the treatment of enteritis, rheumatoid arthritis, organ transplantation and systemic lupus erythematosus. They function by interfering with nucleic acid metabolism in vivo, including de novo and salvage synthetic pathways of nucleic acids. Purine drugs are drugs withou...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N27/447
Inventor 张彦明王玉王燕徐希平于多吴暇
Owner 深圳泰乐德医疗有限公司
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