Primer probe system, method and kit for detecting epidermal growth factor receptor exon 19 and 21 mutations

Abstract

The present invention relates to a primer probe system, a method and a kit for detecting epidermal growth factor receptor exon 19 and 21 mutations. According to the method, an asymmetric PCR technology, a nucleic acid isothermal amplification technology and a fluorescence detection technology are organically integrated, wherein the asymmetric PCR amplification technology and special primers are adopted to obtain a lot of single-stranded DNA so as to overcome disadvantages of low efficiency linear amplification of 3WJ (an isothermal digestion nucleic acid detection technology), and specificity and simplicity of the 3WJ technology are adopted, such that a reaction temperature can be controlled at about 30 DEG C, enzyme activity reduction can not be generated, expensive and precise temperature control equipment is not required during a detection process, and discrimination can be achieved only through the simple fluorescence detection instrument with the special probe design. According to the present invention, advantages of rapidness, simpleness, specificity, sensitivity, and the like are provided.

Description

technical field The invention relates to the technical field of bioengineering, in particular to a primer probe system, method and kit for detecting mutations in exons 19 and 21 of epidermal growth factor receptors. Background technique Lung cancer, especially non-small cell lung cancer, is one of the most malignant tumors in the world. Although traditional chemotherapy has been continuously improved, the prognosis of advanced patients is relatively poor. With the improvement of molecular technology, targeted drugs targeting EGFR (epidermal growth factor receptor) - tyrosine kinase inhibitors (TKI) are becoming more and more prominent in the treatment of NSCLC, among which gefitinib and erythridine Rotinib has been widely used in the treatment of NSCLC. With the accumulation of clinical data, in the treatment of NSCLC, some people respond to TKI treatment. By analyzing the EGFR gene of these TKI-treated patients, it was found that almost all of them had EGFR gene mutation...

AI Technical Summary

Problems solved by technology

Although JP and 3WJ technologies can distinguish SNP polymorphisms well, their linear amplification efficiency and sensitivity are far below the ability to detect DNA in clinical samples, and more importantly, they can only detect single-stranded DNA.

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