BRAF gene mutation detection kit and application thereof

Abstract

The invention discloses a detection kit and detection method for BRAF gene mutations, and belongs to the technical field of B2D10 currently first developed high-tech industrialization important field guide / biology / novel medical precise diagnosis and treatment equipment. The detection kit is characterized by comprising two pairs of specifically amplified primers for BRAF gene mutation loci, an efficient blocking probe of a wild sequence and a BRAF gene specific TaqMan fluorescent probe. The kit can detect specimens of which the number of the mutant copies is as low as 5 to 10, and the mutation content is as low as 0.1%. The detection kit can detect five gene mutations of a BRAF gene simultaneously, the sensitivity is high, operation is easy, detection is low in price, the clinical application range is wide, samples can adopt fresh pathological tissue or paraffin-embedded tissue or a pleural fluid or serum or plasma, the detection speed is high, and only 90 minutes are needed for completing the detection process.

Description

technical field [0001] The invention relates to the fields of biotechnology and medicine, in particular to a rapid detection of BRAF gene mutation related to the selection and diagnosis of tumor medication. 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 China Cancer Registry Annual Report", which shows that there are about ...

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

[0009] 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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