Gene polymorphism detection kit for predicting adverse reaction and curative effect of fluorouracil as well as detection method and application of gene polymorphism detection kit

A technology of genetic polymorphism and detection kits, which is applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of cumbersome operation and time-consuming, and achieve simplified sequencing process and the effect of time

Pending Publication Date: 2021-12-07
菲思特(上海)生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] At present, the methods of obtaining blood DNA mainly include traditional column extraction and magnetic bead extraction, both of which are time-consuming and cumbersome to operate

Method used

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  • Gene polymorphism detection kit for predicting adverse reaction and curative effect of fluorouracil as well as detection method and application of gene polymorphism detection kit
  • Gene polymorphism detection kit for predicting adverse reaction and curative effect of fluorouracil as well as detection method and application of gene polymorphism detection kit
  • Gene polymorphism detection kit for predicting adverse reaction and curative effect of fluorouracil as well as detection method and application of gene polymorphism detection kit

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Experimental program
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Effect test

Embodiment 2

[0067] Embodiment 2, kit detection steps

[0068] The instruments adopted in the present invention are as follows: thermostat, pyrosequencer (Wuhan First Biotechnology Co., Ltd.).

[0069] 1) Take 30 μL EDTA anticoagulated whole blood sample in a PCR amplification tube;

[0070] 2) Add 100 μL of sample treatment solution and 4ul of magnetic beads, and let stand for 5 minutes;

[0071] 3) Place the PCR amplification tube on the magnetic stand, and suck out the mixed solution from the opposite side of the magnetic beads after all the magnetic beads are adsorbed to one side;

[0072] 4) Add amplification reagent 1 into the dry powder of amplification reagent 2, fully dissolve and mix;

[0073] 5) Add 25ul of the prepared PCR reaction solution to the PCR amplification tube obtained in step 4), fully mix the magnetic beads with the PCR reaction solution, centrifuge, and perform a constant temperature reaction;

[0074] 6) Amplify using a PCR instrument, the reaction system is 25...

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Abstract

The invention discloses a gene polymorphism detection kit for predicting adverse reaction and curative effect of fluorouracil as well as a detection method and application of the gene polymorphism detection kit. The detection kit is used for detecting gene polymorphism of fluorouracil metabolism related genes TYMS, GSTP1, NQO1 and MTHFR, the kit designs specific amplification primers and sequencing primers for the TYMS, GSTP1, NQO1 and MTHFR. The kit comprises the following components: a sample treating fluid, magnetic beads, an amplification reagent 1, an amplification reagent 2, the TYMS, the GSTP1, the NQO1, an MTHFR sequencing primer and a positive control. According to the gene polymorphism detection kit for predicting the adverse reaction and the curative effect of the fluorouracil as well as the detection method and application of the gene polymorphism detection kit, rapid DNA preparation, constant-temperature PCR amplification and pyrosequencing technologies are combined to detect the gene polymorphism for predicting the adverse reaction and curative effect of the fluorouracil, and suggestions from the gene perspective are provided for clinical personalized medication.

Description

technical field [0001] The invention relates to a gene polymorphism detection kit for fluorouracil adverse reaction and curative effect prediction, a detection method and application thereof, and belongs to the field of gene detection. Background technique [0002] Fluorouracil drugs are one of the tumor chemotherapy drugs widely used clinically, including 5-fluorouracil, capecitabine, tegafur, carmofur, floxuridine and deoxyfluridine, etc. These drugs are converted into 5-FU, the main pathway is: converted into 5-fluoro-2-deoxyuridine nucleotides, inhibiting thymine nucleotide synthase, blocking the conversion of deoxyuridine nucleotides into deoxythymine nucleotides, thereby Inhibits DNA biosynthesis. In addition, it can also penetrate into RNA in the form of fluorouridine triphosphate (pseudo-metabolite), inhibit RNA synthesis by preventing uracil and orotic acid from penetrating into RNA, and affect protein biosynthesis, thereby inhibiting the proliferation of granulati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/686C12Q1/6869C12N15/11
CPCC12Q1/6886C12Q1/686C12Q1/6869C12Q2600/156C12Q2600/106C12Q2523/308C12Q2521/507C12Q2522/101C12Q2563/143C12Q2563/149C12Q2565/301
Inventor 刘丹孙悦陈立波
Owner 菲思特(上海)生物科技有限公司
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