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A primer, kit and detection method for detecting polymorphism of ugt1a1 gene

A gene polymorphism and kit technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems of incomplete types, long operation time, and difficult interpretation of results, etc., and achieve genetic Comprehensive typing, high sensitivity, and easy operation

Active Publication Date: 2020-12-01
广州迈景基因医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The purpose of the present invention is to solve the problems in the prior art that the types of UGT1A1 gene polymorphism product typing are not comprehensive, the operation time is long, and the results are not easy to interpret, and a kind of primers and kits for UGT1A1 gene polymorphism detection are provided. and detection method

Method used

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  • A primer, kit and detection method for detecting polymorphism of ugt1a1 gene
  • A primer, kit and detection method for detecting polymorphism of ugt1a1 gene
  • A primer, kit and detection method for detecting polymorphism of ugt1a1 gene

Examples

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Embodiment 1

[0054] A primer for UGT1A1 gene polymorphism detection, the primers include primers for UGT1A1 gene promoter region (TA) n repeat typing, primers for UGT1A1*6 (c.211G>A) typing, Primers for UGT1A1*80 (c.-364C>T) typing, where the promoter region (TA) n repeats include UGT1A1*1, UGT1A1*28, UGT1A1*36 and UGT1A1*37;

[0055] Primers for promoter region (TA) n repeat typing of UGT1A1 gene include promoter region (TA) n repeat typing of UGT1A1 gene

[0056] Use the upstream primer and the promoter region (TA) of the UGT1A1 gene to repeat the downstream primer for typing;

[0057] The sequence of the upstream primer for UGT1A1 gene promoter region (TA) n repeat typing is 5'-AAGTACTTTGCTGTGTTCACTCAAG-3';

[0058] The downstream primer sequence for UGT1A1 gene promoter region (TA) n repeat typing is 5'-GTTTCTTTCCTGCCAGAGGTTCGCC-3';

[0059] Primers for UGT1A1*6 (c.211G>A) typing include upstream primers for c.211G typing, upstream primers for c.211A typing and common downstream prim...

Embodiment 2

[0069] A kit for detecting polymorphisms of the UGT1A1 gene, containing the primers in Example 1.

Embodiment 3

[0071] Using the kit in Example 2 to detect the UGT1A1 gene polymorphism detection method comprises the following steps:

[0072] 1) extract the genomic DNA of the sample to be tested;

[0073] 2) The obtained genomic DNA is used as a template, and the PCR amplification reaction is carried out using the kit in Example 2;

[0074] 3) Determine the polymorphism of the UGT1A1 gene according to the results of the PCR amplification reaction.

[0075] The PCR amplification system includes system 1 and system 2, wherein system 1 is used for promoter region (TA) n repeat, c.211G and c.-364C typing amplification, and system 2 is used for c.211A and c.-364T Typing amplification;

[0076] The conditions of the PCR amplification reaction are: 95°C pre-denaturation for 5-15 minutes; 94-95°C, 10-15s, 58-63°C, 30-40s, 72°C, 30s, a total of 35-45 cycles; 72°C, 10min , fully extended;

[0077] The PCR amplification reaction results were analyzed by capillary electrophoresis.

[0078] The ...

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Abstract

The invention discloses a primer and reagent kit for detecting UGT1A1 gene polymorphism. The primer comprises a primer for typing promoter region (TA)n repetition of a UGT1A1 gene, a primer for typingUGT1A1*6 (c.211G>A) and a primer for typing UGT1A1*80 (c.-364C>T), wherein the promoter region (TA)n repetition comprises UGT1A1*1, UGT1A1*28, UGT1A1*36 and UGT1A1*37. The invention also discloses adetection method of the UGT1A1 gene polymorphism. The method is simple and convenient to operate, time-saving and high in flux, the result is easy to discriminate and read, the sensitivity is high, and the genotyping is comprehensive.

Description

technical field [0001] The invention relates to the field of molecular detection, in particular to a primer, a kit and a detection method for detecting UGT1A1 gene polymorphism. Background technique [0002] Irinotecan (CPT-11) is a derivative of semi-synthetic camptothecin, which belongs to an inactive prodrug, and is mainly used for the treatment of advanced / metastatic colorectal cancer in adults, and can also be used for the treatment of small cell lung cancer. treat. The most significant side effects of irinotecan are neutropenia and delayed diarrhea, both of which are dose-limiting side effects, which may lead to death of patients in severe cases. [0003] UGT1A1 is a key enzyme in the metabolic process of irinotecan, so the expression of UGT1A1 gene and its enzyme activity are closely related to the adverse reactions of irinotecan; the detection of UGT1A1 genotype can be used for clinical prediction of serious toxic side effects related to irinotecan, And guide the d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6886C12Q2600/106C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 闫文瑞林威蔡兴盛李梦真邓泱泱孙冠杰杨冬成
Owner 广州迈景基因医学科技有限公司