Probes, primers, detection system and kit for detecting mutations of EGFR gene

Abstract

The invention discloses probes, primers, a detection system and a kit for detecting mutations of an EGFR gene, and the probes and the primers have the sequences of SEQ ID NO.1 to SEQ ID NO.24. The invention is characterized in that (1), the amplification efficiency is greatly improved to a largest extent; (2) the sensitivity is high, and the detection sensitivity can reach 0.2%; (3) compared with a digital PCR method, operations are simple, the cost is saved, and the clinical application scope is wide; (4) plasma samples with large reaction volume are detected, the DNA sampling quantity of the plasma samples becomes larger, the system is more stable, and the detection rate of the plasma samples is increased; (5) 25 mutations of the EGFR gene can be detected simultaneously in three reaction tubes, and results are intuitive and clear; (6) the detection speed is fast, and consumed time is only 1 / 2 that of the digital PCR; and (7) a detection method can detect peripheral blood samples, has convenient sampling, and can dynamically monitor curative effect of EGFR-TKI drugs on patients.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a probe, a primer, a detection system and a kit for detection of epidermal growth factor (EGFR) gene mutation. 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 will cause an increase in the transcription level of genes in the nucleus, resulting in cell proliferation and transformation. Abnormality of EGFR signaling is the cause of many tumors. Epidermal growth factor receptor tyrosine kinase inhibi...

AI Technical Summary

Problems solved by technology

The sequencing method is about 20% less sensitive, and the operation is complicated, the detection time is longer, and the false negative rate is higher

The ARMS-PCR method can achieve a sensitivity of 1%, which can meet the detection requirements for tumor tissue samples, but for blood samples that are convenient to sample, such as circulating tumor cells in plasma or blood, the tumor DNA content is often lower than 1%, ARMS-PCR The sensitivity of the method is not enough to detect the blood, which limits the judgment of clinicians on the efficacy of targeted drugs

In addition, drug-resistant mutations may arise during clinical drug use, and previous methods cannot be detected due to insufficient sensitivity

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