Method of detecting EGFR (epidermal growth factor receptor) gene mutation in urine of colorectal cancer patient by using digital PCR (polymerase chain reaction)

Abstract

The invention discloses a method of detecting EGFR (epidermal growth factor receptor) gene mutation in urine of a colorectal cancer patient by using digital PCR (polymerase chain reaction). The method comprises the particular following steps: extracting a DNA (deoxyribonucleic acid) template from the urine, preparing ddPCR (differential display polymerase chain reaction) liquid, preparing a droplet, performing PCR amplification, detecting the droplet, analyzing data, and displaying a result, wherein a lysate comprises 10mM of Tris-HCl, 1mM of EDTA (ethylene diamine tetraacetic acid) and 0.5% of SDS (sodium dodecyl sulfonate); a thermal circulation mode of detection comprises the steps of incubating at 95 DEG C for 10min, at 95 DEG C for 15s, at 63 DEG C for 1min (45 cycles in total), and finally putting at 4 DEG C. According to the method, ARMS is combined with the digital PCR; the method solves the problems of complication, time consuming, low sensitivity and the like during EGFR gene mutation detection of the colorectal cancer patient in the prior art by detecting the EGFR mutation in the urine of the patient; the non-traumatic detection in addition to pathologic histology is provided particularly.

Description

technical field [0001] The invention relates to the technical field of colorectal cancer detection, in particular to a method for detecting EGFR gene mutation in urine of colorectal cancer patients by applying digital PCR. Background technique [0002] According to the national cancer registration data report of the National Cancer Center, the incidence of colorectal cancer in my country's urban and rural areas ranks third and fifth among all malignant tumors, and the case fatality rate ranks fourth and fifth respectively. The incidence of colorectal cancer is also increasing year by year. Surgery is still the main method for the treatment of colorectal cancer, but the effect is not satisfactory. [0003] Epidermal growth factor receptor (EGFR) EGFR is widely distributed on the cell surface of mammalian epithelial cells, fibroblasts, glial cells, keratinocytes, etc. Play an important role, the loss of protein tyrosine kinase function such as EGFR or the activity of key facto...

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to solve the defects of poor detection accuracy and insufficient sensitivity in the prior art, and provide a method for detecting EGFR gene mutation in the urine of patients with colorectal cancer using digital PCR to solve the above problems

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