Nucleic acid sequence for EGFR gene exon 18 G719X mutation detection, test kit and application of test kit

Abstract

The invention discloses a nucleic acid sequence for EGFR gene exon 18 G719X gene mutation detection, a test kit and application of the test kit, and belongs to the technical field of molecular diagnosis. A primer and probe for detecting the EGFR gene exon 18 G719X mutation, provided by the invention, can be used for specifically amplifying and detecting the G719X mutation of the exon 18 of the EGFR gene. The invention further discloses a detection test kit for detecting the EGFR gene exon 18 G719X gene mutation based on a microdroplet type digital PCR system. The G719X mutation of the EGFR gene exon 18 can be rapidly, sensitively and accurately detected. The kit has the advantages of simplicity and convenience in operation, strong specificity, high sensitivity, accurate result, low cost, high throughput and the like, can be used for rapidly detecting the exon 18 G719X mutation of the EGFR gene clinically, and provides reference basis for clinical diagnosis and treatment of the exon 18G719X mutation of the EGFR gene.

Description

technical field [0001] The invention belongs to the technical field of molecular diagnosis, and in particular relates to a nucleotide sequence, a kit and an application thereof for detecting the mutation of the G719X gene in exon 18 of the EGFR gene. Background technique [0002] The epidermal growth factor receptor gene is located on the short arm of human chromosome 7 and consists of 188,307 bases, including 28 exons. Its tyrosine kinase domain is encoded by exons 18-24. Epidermal growth factor receptor (EGFR) is the expression product of proto-oncogene C-erbB-1 (HER-1), located on the cell membrane. EGFR mainly transmits signals to the nucleus through two pathways, one is Ras→Raf→MAPK pathway; the other is PI3K→PKC→IKK pathway. When the signal is transmitted to the nucleus, it causes an increase in the transcription level of genes in the nucleus, resulting in cell proliferation and transformation. Abnormalities in EGFR signaling are responsible for the development of v...

AI Technical Summary

Problems solved by technology

The method based on the NGS technology platform can detect all possible mutation forms at this site, but the main defect of the NGS method is that the sensitivity is low by about 1%, and the requirements for samples are high and the amount used is large. It can only detect tissue samples, but cannot For tissue sampling, patients who can only collect plasma samples with extremely low DNA content cannot meet the actual detection needs due to low sample concentration and insufficient sensitivity of the methodology, and cannot effectively guide clinical targeted drug use; in addition, the NGS methodological test operation is complicated and the detection process Long, susceptible to interference, poor repeatability, high requirements for technical personnel; difficult to analyze results, requires professional bioinformatics analysts, high cost of instruments and reagents, not suitable for large-scale clinical promotion

The second is the ARMS-PCR method, the sensitivity can only reach 0.5-1% in general, but the detection of plasma samples also has the defect of insufficient sensitivity, which cannot guide clinical targeted drug use; although the operation is relatively simple but the cost is high, the result interpretation needs to use The △Ct format eliminates background interference, negatives are easy to rise, interfere with interpretation, and cause false positives; in addition, the main defect of the ARMS-PCR method is that it can only detect a few mutations, and adding mutation types will lead to non-specific increases, and the detection effect is difficult to guarantee. Therefore, current research and existing products mainly focus on the detection of the most common 2-3 mutations (G719A, G719C or G719A, G719S, G719C). Some mutations cannot be included, which will lead to missed detection of some mutations and affect the targeting of lung cancer patients. medication

At present, many patients cannot successfully collect tumor tissue, and can only use plasma detection, and the existing mainstream methods have the defect of insufficient sensitivity, which will not be able to effectively guide clinical targeted drug use

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