A kit for detecting b-raf gene mutation

Abstract

The invention relates to a B-raf gene mutation detection kit. The B-raf gene mutation detection kit comprises quality-control primer probe internal-standard mixed liquor and detection primer probe internal-standard mixed liquor, wherein the quality-control primer probe internal-standard mixed liquor comprises a quality-control primer pair, a B-raf gene specific probe, an internal-standard primer pair, an internal-standard specific probe and an internal-standard template, and the detection primer probe internal-standard mixed liquor comprises a B-raf gene V600E mutation detection specific primer pair, a B-raf gene specific probe, an amplification blocking nucleic acid sequence, an internal-standard primer pair, an internal-standard specific probe and an internal-standard template. The B-raf gene mutation detection kit has the advantages that an amplification refractory mutation system is combined with a wild type amplification blocking nucleic acid sequence with a phosphorylated terminal, deoxyinosine is introduced into detection of a B-raf gene V600E mutation detection specific ARMS (amplification refractory mutation system) forward primer to enable quality-control PCR (polymerase chain reaction) and detection PCR to be performed for detection of samples parallelly, and each reaction system can have an internal-standard system capable of effectively avoiding false negative and intra-assay variation; the B-raf gene mutation detection kit is low in cost, high in sensitivity and more capable of controlling intra-assay and inter-assay variation.

Description

technical field [0001] The invention relates to a gene mutation detection product, which belongs to the field of biotechnology. Background technique [0002] B-raf is an oncogene, which encodes a serine / threonine-specific kinase, an important transduction factor of the RAS / RAF / MEK / ERK / MAPK pathway, and participates in the regulation of various biological events in cells, such as Cell growth, differentiation and apoptosis, etc. B-raf mutation status is associated with the occurrence and development of various tumors. In a variety of human malignant tumors, such as malignant melanoma, colorectal cancer, lung cancer, thyroid cancer, liver cancer and pancreatic cancer, there are different proportions of B-raf gene mutations, about 66% of malignant melanoma and 15% of colon cancer There is a somatic missense mutation in the B-raf gene. About 80-90% of B-raf gene mutations occur at 1799 nucleotides of exon15, T is mutated to A, resulting in the substitution of glutamic acid enc...

AI Technical Summary

Problems solved by technology

However, the detection cost of this kit is too high, and each specimen needs 3000-5000 yuan, which is not suitable for widespread promotion

2) AmoyDx? BRAFV600E gene mutation detection kit is being produced by Xiamen Aide Biomedical Co., Ltd., which adopts ADx-ARMS? (Specific Primer Double Amplification System) technology, which is a circular primer double amplification technology, and realizes rapid and sensitive detection , but the detection cost is still high

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