Probe, primer and kit for detecting seven kinds of mutations of human NRAS genes

Abstract

The invention discloses a probe, a primer and a kit for detecting seven kinds of mutations of human NRAS genes, wherein in seven groups of primers and probes from NM1 to NM7, the mutation primer in each group can be combined with a conserved sequence of the NRAS genes; the mutation ARMS primers can realize the specific binding with the corresponding mutation sequences; the mutation sequences are amplified; the detection probe can be combined with an amplification segment; the specific binding of the probe with the wild sequence corresponding to the mutation site is blocked; and the wild nonspecific amplification is inhibited. The specific mutation primer and probe blocking technology is used; the seven kinds of mutation types of the NRAS genes can be accurately detected; the built kit for a real-time fluorescent PCR (Polymerase Chain Reaction) amplification reaction system can be used for conveniently and fast detecting the mutations of the NRAS genes; the operation is simple; the result can be easily read; the sensitivity of a detection method is high; 50 copy mutations can be stably detected; the specificity is good; the 20ng wild genome DNA (Deoxyribose Nucleic Acid) does not have specific amplification; and the detection capability is as high as 1 percent.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a probe, primer and kit for detecting seven mutations of human NRAS gene. Background technique [0002] The ras gene family has three genes related to human tumors—HRAS, KRAS and NRAS, which are located on chromosome 11, 12 and 1, respectively. Functionally, Ras protein acts as a molecular switch, and under normal expression, it can regulate cell growth. Abnormal conditions such as point mutation, overexpression or gene translocation can lead to abnormal proliferation of cells, and eventually lead to tumor formation, and more than 30% of human tumors have Ras mutations. As a member of the ras family, NRAS has many similarities with KRAS in structure and function, and with the deepening of research in recent years, it has gradually become another molecular indicator that provides important basis for clinical disease assessment and treatment after KRAS. [0003] NRAS gene mutations a...

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problems of the existing NRAS gene mutation detection methods, such as low detection efficiency, easy sample contamination, long detection time, poor accuracy, and difficulty in discrimination, the present invention provides primers and probes for detecting seven kinds of mutations in human NRAS genes

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