Human EGFR gene mutation site T790M detection reagent and detection method

A technology of T790M and detection reagents, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., which can solve the problems of long cycle, high cost, and difficult clinical promotion, and achieve the effect of trace detection
CN110373467APending Publication Date: 2019-10-25WENZHOU MEDICAL UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
WENZHOU MEDICAL UNIV
Publication Date
2019-10-25

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Abstract

The invention relates to a human EGFR gene mutation site T790Mdetection reagent and detection method. The detection reagent is prepared from a PCR reaction solution I, a PCR reaction solution II, an enzymatic digestion reaction solution,single base extension liquid I, single base extension liquid II and a magnetic particle suspension. The detection method based on the reagent comprises the steps that the PCR reaction solution I and the PCR reaction solution II amplifyan EGFR genein a blood sample, the enzymatic digestion reaction solutionis added for digestion, and the single base extension liquid I, the single base extension solution II and a PCR-SAP product are added into the digested product for single base extension, and product magnetic beads are gathered and then subjected to chip analysis. According to the detection reagent and method used for detecting human EGFR gene p.T790 mutation, as little as 10 mutant DNAs can be detected from 10<4> DNA molecules, the sensitivity reaches0.1%, and T790M micro-detection is achieved.
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Description

technical field

[0001] The invention relates to the technical field of detection of human EGFR gene mutation site p.T790M (c.2369C>T), in particular to PCR technology and single-base extension reaction, combined with magnetic particle technology, to realize the trace detection of p.T790M. Background technique

[0002] EGFR is a glycoprotein receptor on the surface of the cell membrane, has tyrosine kinase (Tyrosine Kinase, TK) activity, and is the expression product of the proto-oncogene c-erbB-1 (HER-1). The main signal transduction pathways of EGFR are: PI3K-PDK pathway, RAS-RAF-MEK-ERK-MAPK pathway, PLC-γ pathway, JAK-STAT pathway. Through these pathways, extracellular signals are converted into intracellular signals, thereby effectively responding to external signal stimuli, regulating cell growth, proliferation, differentiation, and inhibiting cell apoptosis.

[0003] In 2003, the U.S. Food and Drug Administration (FDA) approved AstraZeneca's EGFR small-molecule tar...

Claims

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