Kit and method for detecting gene polymorphism capable of influencing mercaptopurine personalized medications by means of pyro sequencing method

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

Inactive Publication Date: 2012-09-19
周宏灏
View PDF4 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The activity of TPMT varies significantly among patients, 86.6% of the wild-type TPMT*1 popu

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Kit and method for detecting gene polymorphism capable of influencing mercaptopurine personalized medications by means of pyro sequencing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] TPMT (rs1142345) upstream primer: 5′- TGG GGA ATT GAC TGT CTT TTT GA-3′ (SEQ ID NO.2);

[0027] TPMT (rs1142345) downstream primer: 5′- TCC ATT ACA TTT TCA GGC TTT AGC-3′ (SEQ ID NO.3);

[0028] TPMT (rs1142345) sequencing primer: 5′- GAC TGT CTT TTT GAA AAG TT -3′ (SEQ ID NO.4);

[0029] 1. DNA extraction

[0030] 1.1 The preparation and inspection of reagent materials before the experiment are as follows:

[0031] (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.

[0032] 1.2 Take out the EDTA anticoagulant tube containing whole blood from the refrigerator at 4°C, and mix it upside down several times;

[0033] 1.3 Make a mark on the 1.5mL Eppendorf tube corresponding to the unique identi...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a kit and a method for detecting a genetic polymorphism capable of influencing mercaptopurine personalized medications by means of a pyro sequencing method. The genetic polymorphism specifically relates to a thiopurine s-methyltransferase (TPMT) (rs1142345) single nucleotide polymorphism. The kit contains primers shown by SEQID NO.3-6. The kit is capable of achieving accurate, rapid and high throughput detection of the TPMT (rs1142345), so that safe, reasonable and efficient personalized dosing of the mercaptopurine medications can be achieved.

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 polymorphisms of mercaptopurine individualized medication by pyrosequencing. Background technique [0002] Thiopurine is a derivative of 6-mercaptopurine imidazole, which is an antimetabolite with immunosuppressive effect. Alkylation can inhibit the biosynthesis of nucleic acids, prevent cell proliferation, and can cause DNA damage. It affects immunity by suppressing T lymphocytes, which can inhibit delayed allergic reactions. Clinically, it is mainly used for acute and chronic leukemia, choriocarcinoma and organ transplantation. Thiopurine methyltransferase (TPMT) is involved in the methylation of mercaptopurine and azathioprine, and is closely related to its chemotherapy effect and toxicity. The activity of TPMT is affected by genetic polymorphism, which can change the metabolic rate of mercaptopurine through TPMT. The activi...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/68
Inventor 周宏灏
Owner 周宏灏
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap