Digital PCR kit for detecting human EGFR gene mutation

Abstract

The invention provides a digital PCR kit for detecting human EGFR gene mutation, the digital PCR kit is characterized in that a plurality of Ex19Del mutations and L858R mutations are simultaneously detected in a same reaction tube digital PCR reaction system, a mutant type is identified at an Ex19Del site by using a plurality of ARMS primers, and a wild type and a mutant type are identified at an L858R site by using a TaqMan fluorescent probe; after PCR amplification, an Ex19Del mutant type, an L858R mutant type and an L858R wild type are detected through the position and intensity of a fluorescence signal, and absolute quantification is directly carried out on the Ex19Del mutant type, the L858R mutant type and the L858R wild type. The kit is high in flux and low in cost, the Ex19Del25 site and the L858R site are combined into one tube for detection, the detection flux is improved, and the cost of reagent consumables is reduced.

Description

technical field [0001] The invention relates to the technical field of digital PCR, in particular to a digital PCR kit for detecting human EGFR gene mutation. Background technique [0002] In the practice of precision medicine in NSCLC, monitoring the type of gene mutations associated with targeted drugs in patients can be used to evaluate the efficacy. [0003] Epidermal growth factor receptor (EGFR) is an important target of targeted therapy, and its tyrosine kinase domain is encoded by exon 18-24, which plays a crucial role in regulating cell proliferation and differentiation. important role. EGFR gene mutation is a somatic mutation. It is known that human EGFR gene mutations are mainly located in exons 18-21; more than 80% of EGFR mutations in NSCLC patients are deletion mutations in exon 19 (Ex19Del) and L858R mutations in exon 21. Among them, exon 19 deletion mutations accounted for about 46%, while L858R mutations accounted for about 42%. EGFR-TKI drugs have a sig...

AI Technical Summary

Problems solved by technology

This method can only indirectly determine whether there is a sequence inconsistent with the wild type in the deletion region. When there are SNPs or other types of base changes in the deletion region, this method will appear false positives

[0010] 2. Design a dedicated deletion mutation detection probe for each deletion mutant type. This method not only increases the cost, but also causes the total concentration of the probe in the detection system to be too high, resulting in a high fluorescence background value and affecting detection.

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