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Kit and method for detecting gene polymorphism of irinotecan personalized medicine by pyrophosphoric acid sequencing method

A pyrosequencing method and gene polymorphism technology, applied in the field of molecular biology, can solve the problems of increased risk of toxic and side effects of irinotecan, and achieve the effects of simple operation and small sample size

Inactive Publication Date: 2012-08-22
周宏灏
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Problems solved by technology

Studies have shown that UGT1A1*6 mutation reduces the glucuronidation ability of UGT1A1 by 70%, which significantly increases the risk of side effects of irinotecan

Method used

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  • Kit and method for detecting gene polymorphism of irinotecan personalized medicine by pyrophosphoric acid sequencing method
  • Kit and method for detecting gene polymorphism of irinotecan personalized medicine by pyrophosphoric acid sequencing method
  • Kit and method for detecting gene polymorphism of irinotecan personalized medicine by pyrophosphoric acid sequencing method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] UGT1A1*6 (rs4148323) upstream primer: 5′-GAA ATA GTT GTC CTA GCA CC-3′ (SEQ ID NO.3);

[0034] UGT1A1*28 (rs3064744) upstream primer 5'-ACA GCT TTT TAT AGT CAC GTGA-3' (SEQ ID NO.4);

[0035] UGT1A1*6 (rs4148323) downstream primer: 5′-CTT CAA GGT GTA AAA TGC TC-3′ (SEQ ID NO.5);

[0036] UGT1A1*28 (rs3064744) downstream primer 5'-TGC TCA GCC AGT GGC TGC CATCCA-3' (SEQ ID NO.6);

[0037] UGT1A1*6 (rs4148323) sequencing primer: 5′-TCA AGG TGT AAA ATG CTC-3′ (SEQ ID NO.7);

[0038] UGT1A1*28 (rs3064744) sequencing primer: 5'-CGC CCT CTC CTA CTT AT-3' (SEQ ID 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 vari...

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Abstract

The invention discloses a kit and method for detecting gene polymorphism of an irinotecan personalized medicine by a pyrophosphoric acid sequencing method. The kit is used for parting an irinotecan medicine gene, particularly single nucleotide polymorphism of UGT1A1*6 (rs4148323) and UGT1A1*28 (rs3064744). The kit comprises primers shown as SEQ ID NO. 3-8. According to the kit, accurate, quick and high-flux detection of the UGT1A1*6 (rs4148323) and the UGT1A1*28 (rs3064744) can be realized, so that individual administration for realizing safe, rational and effective irinotecan medicine is achieved.

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 gene polymorphism of irinotecan individualized medicine by pyrosequencing. Background technique [0002] Irinotecan is a camptothecin prodrug, which can be metabolized into the active form 7-ethyl-10-hydroxycamptothecin (SN-38) by carboxylesterase in vivo. The target of SN-38 is DNA topoisomerase I, which can interfere with DNA replication and transcription, can inhibit DNA synthesis, and has strong tumor-killing activity. It is widely used in the treatment of solid tumors such as gastric cancer, colorectal cancer, and lung cancer, and can significantly improve the overall survival of patients. However, when irinotecan is used, the incidence of serious side effects is high, especially severe delayed diarrhea and agranulocytosis, which affect its clinical application. Clinical studies have shown that more than 40% of patients receivin...

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 周宏灏
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