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Method for detecting EGFRvIII in tumor tissues

A technology for tumor tissue and tissue samples, applied in the field of molecular biology, can solve the problems of complex operation, cost limitation, and long time consumption, and achieve the effect of short time consumption, high accuracy, and guaranteed accuracy

Inactive Publication Date: 2016-02-03
JINAN UNIVERSITY +2
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have their own advantages, but the complicated operation, long time-consuming, and relatively high cost limit the promotion and use of these methods to a certain extent.

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  • Method for detecting EGFRvIII in tumor tissues
  • Method for detecting EGFRvIII in tumor tissues
  • Method for detecting EGFRvIII in tumor tissues

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Embodiment Construction

[0024] The following will clearly and completely describe the technical solutions of the embodiments of the present invention in conjunction with the accompanying drawings. Apparently, the described embodiments are only some of the embodiments of the present invention, not all of them.

[0025] A method for detecting EGFRvIII in tumor tissue according to an embodiment of the present invention, comprising the following steps:

[0026] 1. EGFRvIII gene sequence analysis and primer design

[0027] Use the written sequence analysis program to scan the gene sequence before the mutation region (71.6% GC content, high repetition rate) base by base to obtain a low-repetition region of 56-70 bases, and use the sequence analysis software to design a specific reaction. The forward primers were matched with the forward primers F56 (SEQ ID NO: 1) and F66 (SEQ ID NO: 3) before the mutation region, and the specificity of each paired primer in the human genome was detected by using the NCBI p...

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Abstract

The invention discloses a method for detecting EGFRvIII in tumor tissues. According to the method, a target gene sequence needing to be detected first undergoes sequence scanning by utilizing a written sequence analysis program, primer selection is performed, so as to design and obtain a nested PCR primer, then tumor tissue RNA extraction and cDNA acquisition are performed, finally a nested PCR method is adopted to perform continuous two-round PCR amplification and then sequencing is performed on an amplification product, and the result is verified through sequence analysis. According to the method, the existence of the EGFRvIII in the malignant tumor tissues can be detected sensitively in a short time, and the good premise and basis are provided for targeted therapy. By adopting the nested PCR method, gene expression signals are amplified through two-round PCR amplification, so that the observed result is simpler, and the applicability of the method is further improved. The amplification product has specificity and guarantees high accuracy and sensitivity in PCR detection.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and specifically relates to a method for detecting EGFRvIII in tumor tissue. Background technique [0002] Malignant tumors are a serious threat to human health and are one of the key public relations diseases in my country and the world, and their incidence is gradually increasing. Oncology research and clinical observations in recent years have found that environmental factors and personal behavior have an important impact on the occurrence and development of human tumors. More and more studies have shown that almost all malignant tumors are related to the environmental factors suffered by the outside world. Various environmental factors cause damage to the genetic material of organisms in different ways, such as gene mutation and recombination, which directly or indirectly cause the activation of proto-oncogenes or the inactivation of tumor suppressor genes, making cells pathologica...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6848
Inventor 李军旗袁晓辉王茜婷何有文廖化新张建
Owner JINAN UNIVERSITY
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