Method and kit for detecting non-small cell lung cancer drive gene mutation spectrum, and application

A non-small cell lung cancer, gene-driven technology, applied in biochemical equipment and methods, microbial determination/examination, etc., can solve the problems of long waiting time for patients, unable to meet the gene mutation spectrum, difficult to obtain materials for advanced patients, etc. Achieve the effect of shortening the detection time, saving the amount of DNA used, and making the results easy

Active Publication Date: 2013-09-18
GUANGDONG GENERAL HOSPITAL
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  • Abstract
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  • Claims
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Problems solved by technology

[0008] At present, the common detection methods of gene mutations include: direct sequencing method, ARMS method, HRM method, DHPLC method, etc. The common disadvantage of these methods is that only one mutation or a mutation of a gene can be detected at a time, and the detection throughput is low, so they can only be detected one by one. Screening for mutation status of driver genes, pa

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  • Method and kit for detecting non-small cell lung cancer drive gene mutation spectrum, and application
  • Method and kit for detecting non-small cell lung cancer drive gene mutation spectrum, and application
  • Method and kit for detecting non-small cell lung cancer drive gene mutation spectrum, and application

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Embodiment 1

[0067] (1) Primer design

[0068] (1) Design primers to amplify the exon segments of the seven driver genes of non-small cell lung cancer

[0069] Primers were used to amplify the exon DNA segments containing hotspot mutation sites in seven driver genes of non-small cell lung cancer, BRAF, EGFR, PIK3CA, PTEN, MEK1, KRAS and NRAS, among which, the exon segment of BRAF BRAF_EX11, BRAF_EX15; the exon segment of EGFR is EGFR_EX18, EGFR_EX19, EGFR_EX20, EGFR_EX21; the exon segment of KRAS is KRAS_EX2; the exon segment of NRAS is NRAS_EX2, NRAS_EX3; the exon segment of PIK3CA PIK3CA_EX9, PIK3CA_EX20; the exon segment of PTEN is PTEN_EX5, PTEN_EX6, PTEN_EX7; the exon segment of MEK1 is MEK1_EX2.

[0070] The designed amplification primers for amplifying the exon segments of the seven driving genes of non-small cell lung cancer are shown in Table 6:

[0071] Table 6 Amplification primers (5'-3') of exon segments of related genes

[0072]

[0073] (2) Design extension primers: de...

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Abstract

The invention discloses a method and a kit for detecting a non-small cell lung cancer drive gene mutation spectrum, and an application. The method comprises the following steps of: designing 15 pairs of amplification primers for amplifying the exon segments of the seven related genes of non-small cell lung cancer, dividing the amplification primers into 6 groups, and preparing an amplification primer mixed solution, performing multiple PCR (polymerase chain reaction) amplification on the to-be-detected samples by the amplification primer mixed solution respectively, and then performing enzymatic digestion; and designing 39 extension primers used for detecting hotspot mutation sites, dividing the extension primers into 6 groups corresponding to the amplification primer mixed solution, and preparing an extension primer mixed solution, performing extension reaction on the digested PCR product, then performing enzymatic digestion, performing capillary electrophoresis on the obtained product, and making a result judgment via software analysis. The kit provided by the invention comprises the amplification primers for amplifying the exon segments of the seven related genes of non-small cell lung cancer, and the extension primers used for detecting hotspot mutation sites. The method and the kit provided by the invention are simple, high in flux, and short in time consumption.

Description

technical field [0001] The invention relates to a method for detecting the mutation spectrum of driving genes of non-small cell lung cancer, in particular to a method, kit and application for detecting the mutation spectrum of driving genes of non-small cell lung cancer. Background technique [0002] The morbidity and mortality of lung cancer rank first among tumors. The traditional treatment method is to select the chemotherapy regimen based on the patient's clinical characteristics and pathological type, but the progression-free period (PFS) is only 6 months. Targeted therapy for lung cancer has made remarkable progress in recent years, the most prominent example being: small molecule tyrosine kinase inhibitors targeting epidermal growth factor (EGFR) in patients with non-small cell lung cancer with mutant EGFR It showed significant efficacy, and the PFS reached 9-13 months. Subsequently, crizotinib targeting the EML4-ALK fusion gene also showed the same clinical efficac...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 吴一龙苏健张绪超安社娟钟文昭
Owner GUANGDONG GENERAL HOSPITAL
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