Primer/probe combination and kit for detecting gene fusion mutation and use method of kit

Abstract

The invention relates to a primer / probe combination and kit for detecting the gene fusion mutation and a use method of the kit. The primer / probe combination comprises at least one, two or three of following components: a primer / probe combination 1 which is used for detecting the gene fusion mutation of an ALK gene and comprises primers 1-8 and probes 1-2; a primer / probe combination 2 which is usedfor detecting the gene fusion mutation of an RET gene and comprises primers 9-13 and a probe 3; and a primer / probe combination 3 which is used for detecting the gene fusion mutation of an ROS1 gene and comprises primers 14-21 and probes 4-6. According to the primer / probe combination, the kit and the use method of the kit, nine fusion mutations of the ALK gene, or four fusion mutations of the RETgene and nine fusion mutations of the ROS1 gene in a cfRNA sample can be simultaneously detected in a single detection hole, and the primer / probe combination, the kit and the use method have the advantages of good specificity, high sensitivity, accuracy in detection and the like.

Description

technical field [0001] The present invention relates to the field of gene detection, in particular to a primer / probe combination, a kit and a use method thereof for detecting gene fusion mutations. Background technique [0002] Lung cancer is a common clinical malignant tumor, and a large number of clinical studies have shown that lung cancer is related to gene changes. Evidence has shown that tyrosine kinases, such as anaplastic lymphoma kinase (ALK), c-ros oncogene 1 receptor tyrosine kinase (c-ros oncogene 1 receptor tyrosine, ROS1) and RET proto-oncogene (RETproto-oncogene, RET) gene fusion phenomenon is related to the diagnosis, prognosis, judgment of treatment effect and development of targeted drugs of lung cancer. It is necessary to detect the gene fusion of the aforementioned tyrosine kinases accurately and efficiently in clinical and scientific research. [0003] cfRNA (circulating cell-free RNA) refers to extracellular free RNA present in body fluids such as ser...

AI Technical Summary

Problems solved by technology

Specifically, first, chromosome banding techniques, fluorescence in situ hybridization, and immunohistochemistry can only be used to detect chromosomes or tissue cells, not cfRNA

Secondly, fluorescent quantitative PCR requires a standard curve to determine the copy number of the sample to be tested, and cannot detect rare mutations in cfRNA; the read length of high-throughput sequencing is short, resulting in inaccurate sequencing results, and the result reporting period is long

Furthermore, the sample abundance of cfRNA is low and the half-life is short, which increases the difficulty of detection. At the same time, ALK involves 9 different gene fusion mutation methods, RET involves 4 different gene fusion mutation methods, and ROS1 involves 9 different gene fusion methods Fusion mutation method further increases the difficulty of detection

Therefore, how to realize the detection of multiple different fusion modes of ALK gene, RET gene or ROS1 gene at the same time, and further how to simultaneously realize the detection of fusion mutations of ALK gene, RET gene and ROS1 gene (22 kinds in total), there are great difficulties

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