A digital pcr kit for detecting mutations in human egfr gene

Abstract

The invention provides a digital PCR kit for detecting human EGFR gene mutations, the digital PCR kit includes simultaneous detection of multiple Ex19Del mutations and L858R mutations in the same reaction tube digital PCR reaction system, wherein the Ex19Del site uses multiple ARMS primers were used to identify the mutant type, and TaqMan fluorescent probe was used to identify the wild type and mutant type at the L858R site; after PCR amplification, the Ex19Del mutant type, L858R mutant type, and L858R wild type were detected by the position and intensity of the fluorescent signal, and compared Absolute quantification is performed directly. The kit of the present invention has high throughput and low cost, and realizes the detection of the Ex19Del25 site and the L858R site in one tube, which improves the detection throughput and reduces the cost of reagent consumables.

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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