Kit used for detecting human KRAS gene mutation and detecting method thereof

Abstract

The invention relates to a kit used for detecting human KRAS gene mutation and a detecting method thereof. The kit comprises a mutation detecting reaction primer and an inner reference reaction primer. The kit used for detecting the human KRAS gene mutation disclosed by the invention is designed based on an ARMS-PCR experimental method, is simple and reasonable in structural design and is convenient to use, and has the advantages of being high in sensitivity, strong in specificity, short in time consumption and the like while being used for detecting KRAS gene mutation.

Description

technical field [0001] The invention relates to a kit for detecting human KRAS gene mutation and a detection method thereof, belonging to the field of molecular biology. Background technique [0002] 1) Clinical significance of KRAS gene mutation in colorectal cancer [0003] In recent years, with the continuous improvement of people's living standards, changes in eating habits and dietary structure, and population aging, the incidence and mortality of colorectal cancer (CRC) in my country have maintained an upward trend. Among them, the incidence of colon cancer increased significantly. Most patients are already in the middle and late stages when they are discovered. The incidence of colorectal cancer is 221,000, the mortality rate is 110,000, and the 5-year survival rate is 63.7%. [0004] KRAS is a key downstream regulator in the EGFR (epidermal growth factor receptor) signaling pathway, which is involved in the regulation of cell growth and plays an important role in ...

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Problems solved by technology

[0011] Three EGFR-TKIs (gefitinib / Iressa, erlotinib / Tarceva, and Camino) are currently the main drugs for molecularly targeted therapy in NSCLC, but in different patients EGFR- Efficacy of TKIs varies widely

Among them, the sanger sequencing method can directly obtain the mutation information of the sample through sequencing, but this method requires the sample to be tumor tissue, and the sensitivity is not high. It can only detect mutations with a mutation frequency of more than 20%, which is easily affected by background DNA. For other samples than tumor tissue

In addition to the direct sequencing method, the PCR-SSCP and PCR-RFLP methods are only used for the detection of tumor tissues, and there is no introduction of detection in other specimens

High-throughput sequencing takes a long time and costs a lot

Digital PCR has high sensitivity, but low throughput and high cost

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