Method and kit for detecting colorectal cancer KRAS/NRAS/BRAF/PIK3CA genes

Abstract

The invention relates to the field of biotechnology, in particular to a method and kit for detecting colorectal cancer KRAS / NRAS / BRAF / PIK3CA genes based on a high-throughput sequencing platform. The method comprises the steps of extraction of sample DNA, amplification of a target fragment, digestion of a primer sequence, connection of joints, amplification and purification of a library, quality testing of the library, template preparation and enrichment, online sequencing and data analysis. By the adoption of the method and kit for detecting colorectal cancer KRAS / NRAS / BRAF / PIK3CA genes, detection efficiency, accuracy and sensitivity are greatly improved, detection cost is reduced, and a more definite basis can be provided for individualized treatment of colorectal cancer patients.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method and a reagent for detecting colorectal cancer KRAS / NRAS / BRAF / PIK3CA genes based on an Ion Proton sequencing platform. Background technique [0002] As the most common malignant tumor of the digestive tract, colorectal cancer ranks third in incidence and fourth in mortality in the world. There are more than one million new cases of colorectal cancer every year in the world, and the incidence rate ranks third in male malignant tumors and second in female malignant tumors. The morbidity and mortality of colorectal cancer are higher in economically developed countries and regions than in less developed countries and regions, and the geographical distribution of male and female incidence is consistent, but males are higher than females. The risk of colorectal cancer morbidity and mortality increases with age. The global morbidity and mortality of colorectal cancer are still on ...

AI Technical Summary

Problems solved by technology

The PCR method detects mutation points. Usually, only one mutation site can be detected in one system, which is prone to false positives; the detection of multiple mutation sites is cumbersome and requires a large sample size

The first-generation sequencing method can only sequence a certain region of a sample at a time, so the detection efficiency is low and the detection cost is high; and due to the limitation of the sequencing principle of the first-generation sequencing method, double peaks will appear when detecting the signal of a sample with a heterozygote Even multi-peak, so that the sample can not be accurately identified, resulting in low detection sensitivity (20%), high false negative rate

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