EGFR gene detection kite and detection method

A gene detection and kit technology, which is applied in the fields of genetic engineering and molecular biology, can solve the problems of low sensitivity and low accuracy of EGFR detection, and achieve the effect of low detection false positive rate and high sensitivity

Inactive Publication Date: 2018-04-06
GUANGZHOU KANGXINRUI GENE HEALTH TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The object of the present invention is to provide a EGFR gene detection kit and detection metho

Method used

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  • EGFR gene detection kite and detection method
  • EGFR gene detection kite and detection method
  • EGFR gene detection kite and detection method

Examples

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Effect test

no. 1 example

[0027] The present invention proposes a first embodiment, an EGFR gene detection kit, including a primer set for specific amplification of at least one of a plurality of EGFR gene test sites, the test site includes rs28929495, rs121913428、rs121913421、rs397509368、rs121913442、rs121913438、rs397517098、rs727504339、rs121913435、rs121913440、rs121913436、rs727504258、rs727504233、rs121913422、rs727503016、rs727503015、rs727504332、rs121913465、rs121434569、rs121434568中的至少一个;用于特异性扩增含rs28929495 And / or the primer set of the fragment of rs121913428 site comprises SEQ ID NO:1 and SEQ ID NO:2, SEQ ID NO:3 and SEQ ID NO:4, SEQ ID NO:1 and SEQ ID NO:4, SEQ ID NO:1 At least one pair of NO:3 and SEQ ID NO:2;

[0028] 用于特异性扩增含rs121913421、rs397509368、rs121913442、rs121913438、rs397517098、rs727504339、rs121913435、rs121913440、rs121913436、rs727504258、rs727504233、rs121913422、rs727503016、rs727503015、rs727504332中至少一个位点的片段的引物组包括SEQ ID NO At least one pair of: 5 and SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, SEQ I...

no. 2 example

[0041] The present invention provides a second embodiment, a method for detecting EGFR gene, comprising the following steps:

[0042] A. Amplify at least one EGFR nucleic acid fragment containing the site to be tested to obtain an amplified product containing the site to be tested; the site to be tested includes rs28929495, rs121913428, rs121913421, rs397509368, rs121913442, rs121913438, rs397517098, rs727504339 、rs121913435、rs121913440、rs121913436、rs727504258、rs727504233、rs121913422、rs727503016、rs727503015、rs727504332、rs121913465、rs121434569、rs121434568中的至少一个;用于特异性扩增含rs28929495和 / 或rs121913428片段的引物组包括SEQ ID NO: 1 and at least one pair of SEQ ID NO:2, SEQ ID NO:3 and SEQ ID NO:4, SEQ ID NO:1 and SEQ ID NO:4, SEQ ID NO:3 and SEQ ID NO:2; 于特异性扩增含rs121913421、rs397509368、rs121913442、rs121913438、rs397517098、rs727504339、rs121913435、rs121913440、rs121913436、rs727504258、rs727504233、rs121913422、rs727503016、rs727503015、rs727504332中至少一个位点的片段的引物组包括SEQ ID NO: 5 and at least one pair of SEQ ID...

no. 3 example

[0055] The present invention provides a third embodiment, an EGFR gene detection method, using human whole blood genome sample 1 and human whole blood genome sample 2 as templates, and respectively establishing a reaction system according to the following steps:

[0056] A. For the above two samples, prepare a nucleic acid fragment for amplifying EGFR exon 18, a nucleic acid fragment for EGFR exon 19, two nucleic acid fragments for EGFR exon 20, and a nucleic acid fragment for EGFR exon 21 PCR amplification reaction systems for regional nucleic acid fragments, a total of 2 groups, 10 systems; configure each reaction system according to the following ratio: 10ng / μL human whole blood DNA molecule 5.0μL; 2×long Taq Mix (Shenzhen Huayinkang Gene Technology Co., Ltd.) 10 μL; 0.5 μmol / mL upstream primer 0.5 μL; 0.5 μmol / mL downstream primer 0.5 μL; 4 μL deionized water; mix well and centrifuge. Then place the centrifuge tube in the PCR machine and set the reaction program: 4 minutes...

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Abstract

The invention relates to the fields of genetic engineering and molecular biology and provides an EGFR gene detection kit and a detection method. The kit is characterized in that a specific amplification primer sequence is designed specific to specific sites of EGFR No.18 exon, No.19 exon, No.20 exon and No.21 exon regions, so that detection false positive rate of a to-be-detected site is low, sensitivity is high, and genetic mutation with mutation ratio higher than or equal to 5% can be detected; an F joint and an R joint are respectively connected with the two ends of an EGFR nucleic acid fragment containing the to-be-detected site to obtain library molecules, the F joint and the R joint respectively contain an amplification primer binding site, and when sequencing is carried out on multiple EGFR gene fragments containing to the to-be-detected site, an amplification primer used for amplifying the library molecules is unified, so that amplification efficiency consistency is guaranteed,and primer design difficulty is reduced; and the R joint is designed to be of a hairpin structure, and the tail end of an amplified product can be closed, so that the F joint is effectively avoided from being connected on the R joint, and connecting efficiency is improved.

Description

technical field [0001] The invention relates to the fields of genetic engineering and molecular biology, and more specifically relates to an EGFR gene detection kit. Background technique [0002] Epidermal growth factor receptor (EGFR) is a member of the cell surface receptor tyrosine kinase HER / ErbB family, widely distributed on the cell membrane of various normal human tissues, and plays an important role in cell proliferation and migration. effect. Tyrosine kinase inhibitors (TKIs) are drugs targeting EGFR. The most important factor affecting the efficacy of TKIs is the mutation of the EGFR gene. The mutations associated with the efficacy of TKIs are mainly located in the EGFR gene. On exons 18, 19, 20, and 21. Therefore, the detection of EGFR gene mutation has important reference value for guiding patients' clinical medication. [0003] At present, the commonly used detection methods of EGFR gene mutation mainly include Sanger method, real-time fluorescent quantitativ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6869
CPCC12Q1/6869C12Q1/6888C12Q2600/106C12Q2600/156C12Q2525/191C12Q2535/122C12Q2521/301C12Q2521/501
Inventor 盛司潼高虹
Owner GUANGZHOU KANGXINRUI GENE HEALTH TECH CO LTD
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