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A gene detection kit, primer and method for lung cancer targeted drug

A gene detection and kit technology, which is applied in the field of gene detection kits for lung cancer targeted drugs, can solve the problems of specificity, accuracy, low uniformity, and difficulty in implementing multiple PCR capture technology, and achieve ideal sensitivity results

Active Publication Date: 2020-01-07
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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  • A gene detection kit, primer and method for lung cancer targeted drug
  • A gene detection kit, primer and method for lung cancer targeted drug
  • A gene detection kit, primer and method for lung cancer targeted drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1. Specific primer set for detection of lung cancer targeted drug genes

[0021] The inventor designed specific primers according to 28 target regions of lung cancer targeted drug-related genes, and screened, optimized, and verified through a large number of tests, and finally selected 28 pairs of primers with high amplification efficiency and good specificity (as shown in Table 1). Show).

[0022] Table 1. Specific primers for lung cancer targeted drug genes

[0023]

[0024]

[0025] The 28 target regions amplified by the specific primers were used to detect the mutation sites related to the lung cancer targeted drug genes shown in Table 2.

[0026] Table 2. Mutation sites related to lung cancer targeted drug genes

[0027]

[0028]

[0029]

[0030]

[0031]

[0032]

Embodiment 2

[0033] Example 2, lung cancer targeted drug gene detection kit

[0034] According to the semiconductor sequencing platform, this embodiment exemplarily provides a lung cancer targeted drug gene detection kit based on the semiconductor sequencing method. The gene detection kit for lung cancer targeted drugs based on sequencing should also be within the scope of protection of this application, and will not be exemplified here.

[0035] Gene detection kits for lung cancer targeted drugs, including:

[0036] (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:56, and the common primer sequence is: 5'-AAATGGGCGGTAGGCTTG-3 '(SEQ ID NO: 57);

[0037] (2) Sequencing adapter: 5'-CCTCTCTATGGGCAGTCGGTGAT-common primer-3' (SEQ ID NO:58);

[0038] (3) Index adapter: 5'-CCATCTCATCCCTGCGTGTCTCCGACTCAGNNNNNNNNNGAT-common primer-3' (SEQ ID NO: 59); N in the sequence represents any base of AGCT, and NNNNNNNNN is used to identify l...

Embodiment 3

[0041] Example 3. A method for library construction and sequencing of lung cancer targeted drug genes

[0042] This embodiment exemplarily provides a method for library construction and sequencing of lung cancer targeted drug genes, the steps are as follows:

[0043] 1. The first step of PCR reaction and purification

[0044] Prepare 25 μL of the first step PCR reaction system, including: 10-20 ng of template DNA, 2 μL of specific primer set, 12.5 μL of amplification buffer, and the rest of nuclease-free water.

[0045] Amplification was carried out according to the following procedure: ①95°C for 3 min; ②15 cycles of amplification, each cycle: 95°C for 20 s, 60°C for 90 s, 72°C for 30 s; ③72°C for 1 min, ④16°C for maintenance.

[0046] The PCR product of the first step was purified using a purification reagent to obtain 11.5 μL of the purified PCR product of the first step.

[0047] 2. The second step of PCR reaction

[0048] Prepare 25 μL of the second-step PCR reaction sy...

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Abstract

The invention discloses a detection kit, primer and method for a targeted drug administration gene of lung cancer. A specific primer group is designed aiming at 28 target regions of the targeted drugadministration gene of the lung cancer, and the detection kit for the targeted drug administration gene of the lung cancer is provided; and the kit and the specific primer group can be utilized for rapidly and accurately detecting hundreds of mutation sites of the targeted drug administration gene of the lung cancer once, the uniformity, the repeatability, the library-building success rate, the utilization rate of library sequencing data, the detection accuracy and the sensitivity are very ideal in a multi-PCR library-building process through a specific primer, and the kit and the specific primer group are suitable for multiple sample types.

Description

technical field [0001] The invention belongs to the field of gene detection, and more specifically relates to a lung cancer targeted drug gene detection kit, primers and method. Background technique [0002] Lung cancer is the main cause of cancer death. According to statistics, in 2015, there were 7.333 million lung cancer cases and 6.102 million deaths in the Chinese population, both ranking first. Genotyping can guide the diagnosis and treatment of lung cancer, and gene testing to guide the treatment of lung cancer has become a new medical model. For example, the National Comprehensive Cancer Network (NCCN) of the United States issued guidelines stating that when diagnosing and treating non-small cell lung cancer, EGFR, ALK and other gene tests are required; when EGFR has sensitive mutations, EGFR-TKIs targeted drugs are recommended as first-line drugs; KRAS and other mutations exist EGFR-TKIs targeted drugs are not recommended. In addition, studies have shown that the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6886C12Q2600/106C12Q2600/156C12Q2600/16C12Q2537/143
Inventor 黄铨飞倪晨子吴春求韩冉冉吕来灰李慧
Owner CAPITALBIO GENOMICS
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