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Detection kit for daunorubicin metabolism marker and detection method and application thereof

A technology of metabolic markers and detection kits, applied in the field of gene detection, can solve the problems of complicated operation steps, easy pollution and low specificity, etc.

Pending Publication Date: 2021-11-19
菲思特(上海)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the sequencing method and chip method have cumbersome operation steps, long detection cycle, and the amplification products are prone to contamination; the high-resolution melting curve method, allele-specific amplification method, and Taqman fluorescent probe method have simple steps and high specificity. Low, and high requirements for instruments and equipment or high test costs

Method used

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  • Detection kit for daunorubicin metabolism marker and detection method and application thereof
  • Detection kit for daunorubicin metabolism marker and detection method and application thereof
  • Detection kit for daunorubicin metabolism marker and detection method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1, the preparation of kit

[0044] The kit of the present invention designs specific amplification primers and sequencing primers for SLC28A3 (C1381T) and UGT1A6*4 (G>T) for amplification and pyrosequencing detection. Designing primers based on rapid amplification technology is one of the keys of the present invention. Gene polymorphism sequence is subject to the public sequence in Genebank.

[0045] (1) The primer sequences of this embodiment are as follows:

[0046]

[0047]

[0048] (2) The detection kit of the present embodiment comprises the following components:

[0049] serial number Component Marked capacity 1 Amplification primers, blood sample direct PCR master mix, trehalose 400μL×1 tube 2 SLC28A3(C1381T) Sequencing Primers 70ul×1 tube 3 UGT1A6*4(G>T) sequencing primer 70ul×1 tube 4 positive control 50μL×1 tube

[0050] (3) The single serving configuration system of the detection kit PCR re...

Embodiment 2

[0054] Embodiment 2, pyrophosphate detection

[0055] The instruments adopted in the present invention are as follows: amplification instrument, pyrosequencer (Wuhan First Biotechnology Co., Ltd.).

[0056] (1) Reagent preparation (reagent preparation room)

[0057] Take out the reagents in advance, vortex the PCR reaction solution for 15 seconds, and centrifuge at low speed for later use. Determine the number of reactions N, N = number of samples to be tested (n) + number of quality control products (1) + blank control. It is recommended to conduct positive control and blank control analysis for each PCR experiment at the same time. Then the reaction solution was dispensed into PCR reaction tubes at 16 μL / tube.

[0058] (2) Sample testing (sample preparation room)

[0059] Add EDTA anticoagulated whole blood, positive control and blank control into the PCR reaction tube according to the sample volume of 4 μL, close the tube cap tightly, centrifuge at low speed for 15 seco...

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Abstract

The invention discloses a detection kit for daunorubicin metabolic markers and a detection method and application thereof, the detection kit is used for detecting the polymorphism of two genes SLC28A3 and UGT1A6 * 4 of the daunorubicin metabolic markers, the kit designs a specific amplification primer and a sequencing primer according to the polymorphism of the two genes SLC28A3 and UGT1A6 * 4; and the kit comprises the following components: an amplification reaction solution, an SLC28A3 sequencing primer, a UGT1A6 * 4 sequencing primer and a positive control. According to the invention, blood direct expansion, rapid amplification and optimized pyrosequencing technologies are combined to detect the gene polymorphism related to daunorubicin curative effect and adverse reaction prediction, and suggestions from the gene perspective are provided for daunorubicin clinical personalized medication.

Description

technical field [0001] The invention relates to a detection kit for daunorubicin metabolic markers, a detection method and application thereof, and belongs to the field of gene detection. Background technique [0002] Anthracyclines are chemical substances with anti-tumor activity produced by microorganisms, including doxorubicin (doxorubicin), epirubicin (erubicin), daunorubicin and aclarmycin, etc. It is widely used in the treatment of hematological malignancies and solid tumors, including acute leukemia, lymphoma, breast cancer, ovarian cancer, gastric cancer and soft tissue sarcoma, etc. Its mechanism of action is to insert DNA molecules non-specifically between adjacent base pairs , combined with the ribose-phosphate backbone, resulting in partial unwinding of the DNA molecule, and can interfere with topoisomerase II to reconnect the broken DNA double strands, thus hindering the process of DNA replication, transcription and RNA synthesis. In the body, anthracyclines ar...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6858C12N15/11
CPCC12Q1/6886C12Q1/6858C12Q2600/106C12Q2600/156C12Q2600/166C12Q2600/16C12Q2531/113C12Q2537/143C12Q2535/122C12Q2545/113
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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