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Individualized gene detection kit, primers and method for tumor chemotherapeutic drugs

A gene detection and chemotherapeutic drug technology, which is applied in the field of individualized gene detection kits for tumor chemotherapeutic drugs, can solve the problems of difficulty in implementing multiple PCR capture technology, low specificity, accuracy, and low uniformity, and achieve ideal detection accuracy Effect

Active Publication Date: 2019-07-12
CAPITALBIO GENOMICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, generally speaking, the more target regions to be captured by multiplex PCR, the lower the specificity, accuracy, and uniformity of amplification, and the more difficult it is to realize the multiplex PCR capture technology

Method used

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  • Individualized gene detection kit, primers and method for tumor chemotherapeutic drugs
  • Individualized gene detection kit, primers and method for tumor chemotherapeutic drugs
  • Individualized gene detection kit, primers and method for tumor chemotherapeutic drugs

Examples

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Effect test

Embodiment 1

[0027] Example 1. Specific primer set for detecting individualized gene of tumor chemotherapeutics

[0028] The inventor has designed specific primers according to 13 polymorphic sites of individualized genes of tumor chemotherapeutic drugs, and screened, optimized, and verified through a large number of experiments, and finally selected 13 pairs of primers with high amplification efficiency and good specificity (such as shown in Table 1).

[0029] Table 1. Specific primers for personalized genes of tumor chemotherapy drugs

[0030]

[0031]

Embodiment 2

[0032] Embodiment 2, tumor chemotherapeutic drug individualized gene detection kit

[0033] According to the semiconductor sequencing platform, this embodiment exemplarily provides an individualized gene detection kit for tumor chemotherapeutics based on the semiconductor sequencing method. Non-sequencing-based personalized gene detection kits for tumor chemotherapeutics should also be within the protection scope of this application, and will not be exemplified here.

[0034] Personalized gene detection kits for tumor chemotherapy drugs, including:

[0035] (1) Specific primer set: Add a common primer to the 5' end of each specific primer shown in SEQ ID NO:1~SEQ ID NO:26, and the common primer sequence is: 5'-AAATGGGCGGTAGGCTTG-3 '(SEQ ID NO::27);

[0036] (2) Sequencing adapter: 5'-CCTCTCTATGGGCAGTCGGTGAT-common primer-3' (SEQ ID NO: 28);

[0037] (3) Index adapter: 5'-CCATCTCATCCCTGCGTGTCTCCGACTCAGNNNNNNNNGAT-common primer-3' (SEQ ID NO: 29); N in the sequence represents...

Embodiment 3

[0040] Example 3. Library construction and sequencing of personalized genes for tumor chemotherapy drugs

[0041] This embodiment exemplarily provides a library construction and sequencing method for individualized genes of tumor chemotherapeutic drugs, the steps are as follows:

[0042] 1. One-step multiplex PCR

[0043] Prepare a 25 μL multiplex PCR reaction system, including: template DNA 300ng, specific primer set 10pM, sequencing adapter 10pM, index adapter 10pM, amplification buffer 12.5μL, nuclease-free water balance; different templates use different index adapters.

[0044] Amplification was performed according to the following procedure: ① 95°C for 3 min; ② 35 cycles of amplification, each cycle: 95°C for 10 s, 60°C for 90 s, 72°C for 30 s; ③ 72°C for 1 min, and ④ 16°C to hold.

[0045] 2. Amplified product mixing and purification

[0046]Take out 10 μL of the amplified product from each PCR tube, vortex and mix to obtain a mixed library; take 25 μL of the mixed li...

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Abstract

The invention discloses an individualized gene detection kit, primers and a method for tumor chemotherapeutic drugs. The specific primer group is designed for 13 polymorphic loci of individualized genes of the tumor chemotherapeutic drugs, and provided is the individualized gene detection kit for the tumor chemotherapeutic drugs. The kit and the specific primer group can detect 13 individualized gene polymorphic loci of the tumor chemotherapeutic drugs quickly and accurately at a time. Moreover, the uniformity, repeatability, library building success rate, the utilization rate of library sequencing data and the detection accuracy of multiplex PCR library building by the specific primers are ideal.

Description

technical field [0001] The invention belongs to the field of gene detection, and more specifically relates to a kit, primer and method for individualized gene detection of tumor chemotherapeutic drugs. Background technique [0002] There are individual differences in drug effects, and its influencing factors include physiological factors, pathological factors, environmental factors and genetic factors, among which genetic factors mainly depend on individual gene polymorphisms, including single nucleotide polymorphisms and copy number polymorphisms , these polymorphisms will affect pharmacokinetics and pharmacodynamics through drug metabolizing enzymes, drug transporters, drug receptors, pathogenic genes, etc., resulting in individual differences in drug efficacy and toxicity. The traditional medical model is to formulate a treatment plan according to the patient's condition, and the treatment result may appear in three situations: effective, ineffective, and adverse reaction...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11C40B50/06C40B40/06
CPCC12Q1/6886C12Q2600/106C12Q2600/156C40B40/06C40B50/06
Inventor 黄铨飞韩冉冉吴春求倪晨子吕来灰李慧
Owner CAPITALBIO GENOMICS