Fluorescent genotyping detection kit and detection method for eight K-ras gene mutations

Abstract

The invention belongs to the field of biotechnology and clinical molecular diagnosis and in particular relates to a fluorescent genotyping detection kit and a detection method for eight human K-ras gene mutations. According to the method, an MGB (minor groove binder) probe technology is combined with a mismatch ARMS (amplification refractory mutation system) technology, an MGB probe complementary with a DNA (deoxyribonucleic acid) plus strand and an ARMS downstream primer complementary with a DNA minus strand are designed at allelic gene loci, a mismatch locus is introduced in the ARMS downstream primer, the MGB probe is overlapped with the ARMS primer by not more than 5 bp, and the eight human K-ras gene mutations are subjected to genotyping detection by four reaction pipes based on multiple fluorescent PCR (polymerase chain reaction). The method has the advantages of good specificity, high sensitivity, simplicity and quickness for operation, accuracy for genotyping, simplicity for result interpretation and the like, and can be used for detecting the clinical K-ras gene mutations and helping doctors screen the crowed with effective anti-EGFR (epidermal growth factor receptor) treatment.

Description

technical field [0001] The invention belongs to the fields of biotechnology and clinical molecular diagnosis, and in particular relates to a detection kit and a detection method for eight mutations of human K-ras gene. Background technique [0002] K-ras gene is one of the RAS gene family, located on human chromosome 12, is a proto-oncogene, which has a great impact on human cancer. The K-ras gene encodes a ras protein with a molecular weight of 21kD, also known as p21 protein. The p21 protein is located on the inner surface of the cell membrane, has GTPase activity, and participates in intracellular signal transduction. It is active when combined with GTP, and inactive when combined with GDP. . The K-ras gene is like a molecular switch in the body. It plays an important regulatory role in the signal transduction pathway of tumor cell growth and angiogenesis. The normal K-ras gene can inhibit the growth of tumor cells, but it will induce The gene is permanently activated, ...

AI Technical Summary

Problems solved by technology

At present, the domestic Xiamen Aide Biomedical Technology Co., Ltd. has developed the ADx-ARMS patented technology with independent intellectual property rights on the basis of ARMS technology, and successfully developed the corresponding kit for K-ras gene mutation detection, but the company The kit is expensive, and the detection of 7 mutations needs to be detected in 7 tubes, requiring a large amount of tumor samples