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Probe and primer combination for detecting L861Q locus mutation of human EGFR gene

A technology of L861Q and primer combination, which is applied in the fields of biotechnology and medicine, can solve the problems of strong background, time-consuming, low sensitivity, etc., and achieve the effects of simple and fast operation, simple result interpretation, and high detection sensitivity

Inactive Publication Date: 2016-03-09
SOUTH CENTRAL UNIVERSITY FOR NATIONALITIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the main method for point mutation detection is sequencing, which is time-consuming and has low sensitivity.
Other fluorescent quantitative PCR methods used in clinical detection have relatively high sensitivity, but the background is strong, which is prone to false positive results

Method used

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  • Probe and primer combination for detecting L861Q locus mutation of human EGFR gene
  • Probe and primer combination for detecting L861Q locus mutation of human EGFR gene
  • Probe and primer combination for detecting L861Q locus mutation of human EGFR gene

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Experimental program
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Effect test

Embodiment 1

[0046] Firstly, the EGFR gene L861Q site mutation detection system provided by the present invention is prepared. Include the following steps:

[0047] 1. Primer and probe synthesis

[0048] Synthetic primer sequences SEQIDNO:1, SEQIDNO:2, SEQIDNO:4, SEQIDNO:5, SEQIDNO:7, SEQIDNO:8; Synthetic specific probe sequences SEQIDNO:3, SEQIDNO:6, SEQIDNO:9, and at the 5' end Label the FAM or HEX fluorescent group, and connect the BHQ1 modification group to the 3' end.

[0049] The above primers were respectively prepared into 100 μM mother solution for storage, and the above probes were respectively prepared into 100 μM mother solution for storage.

[0050] 2. Preparation of fluorescent quantitative PCR reaction system

[0051] The quality control detection reaction system and the mutation detection reaction system were prepared respectively, and the components are shown in the following table:

[0052]

[0053] The method of the present invention is used to detect the mutation...

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Abstract

The invention is suitable for the field of the biotechnology and medicine, and provides a method for detecting L861Q locus mutation of the human EGFR gene. Particularly, the fluorescent quantitative PCR technology, an ARMS primer and the TaqMan fluorescent probe technology are combined, and the method for detecting L861Q locus mutation of the human EGFR gene is invented. The method involves a specific primer and a specific probe. The method has the advantages of being simple, effective, high in sensitivity, high in specificity and easy and rapid to operate.

Description

technical field [0001] The invention provides a method for detecting the mutation of the L861Q site of the human EGFR gene, and provides related amplification primers and probes, belonging to the fields of biotechnology and medicine. Background technique [0002] EGFR (Epidermal Growth Factor Receptor, also known as: Epidermal Growth Factor Receptor) gene, a member of the Epidermal Growth Factor Receptor (HER) family, plays an important regulatory role in cell physiological processes. The EGFR gene is located in the 7p12-14 region of the short arm of chromosome 7. It consists of 118kb and contains 28 exons. Among them, the tyrosine kinase functional region is encoded by exons 18-24, and exons 18-20 encode N Telolobule, exons 21-24 encode the C-terminal lobe. EGFR protein is a transmembrane protein with receptor tyrosine kinase activity, present on the surface of mammalian epithelial cells, keratinocytes, glial cells, fibroblasts and other cells. EGFR and HER2 (erbB2), HER3...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6858C12Q2563/107C12Q2545/101
Inventor 沈金花黄俊
Owner SOUTH CENTRAL UNIVERSITY FOR NATIONALITIES