Kit for detecting EGFR gene mutation and application of kit

Abstract

The invention discloses a kit for detecting EGFR gene mutation and an application of the kit. The kit is characterized by containing 20 specific amplification primers for an EGFR gene mutation site, 5 efficient blocking probes for wild sequences, and 4 EGFR gene specific TaqMan fluorescent probes. The kit can be used for detecting samples of which the mutation copy number is as low as 5-10 copies and the mutation content is as low as 0.1%. The kit disclosed by the invention can be used for simultaneously detecting 5 types of gene mutations of an EGFR gene, is high in sensitivity, simple to operate, low-cost in detection and wide in clinical application range, the samples can be fresh pathological tissues, paraffin embedded tissues, pleural fluid, serum or plasma, the detection speed is high, and the detection process can be finished in only 90 minutes.

Description

technical field [0001] The invention relates to an EGFR gene mutation detection kit and application thereof, in particular to a rapid detection of EGFR gene mutation related to selection and diagnosis of tumor medication. Belongs to the field of biotechnology and medicine. Background technique [0002] The World Health Organization published the "World Cancer Report" on the occasion of World Cancer Day in 2014, stating that cancer has become the leading cause of death in the world, and both morbidity and mortality are on the rise. The global cancer incidence rate has increased by 11% in four years, and by 2012 there were approximately 14.1 million cases and 8.2 million deaths due to cancer. Half of the new cases of cancer worldwide in 2012 occurred in Asia, most of them in mainland China. And predicted that in the next 20 years, the world's cancer cases will increase by 75%, reaching nearly 25 million. In early 2013, the China Cancer Registry Center released the "2012 Chi...

AI Technical Summary

Problems solved by technology

The direct sequencing method has always been considered the gold standard for detecting gene mutations. This method is relatively cheap, but it takes a long time to operate, and it has two obvious disadvantages: one is low sensitivity, and generally requires the abundance of mutated genes to reach more than 20%. in order to accurately detect

Second, due to the subsequent need to process the PCR product, contamination is prone to occur and the results are inaccurate

[0008] However, due to the low content of circulating DNA in peripheral blood, the content of tumor-specific mutant DNA is even less, especially in early cancer patients. In the same amplification system, a large number of wild-type templates are dominantly amplified, resulting in the inhibition of the amplification of a small amount of mutant templates and cannot be detected

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