Primer combination for detecting human EGFR, KRAS and BRAF gene mutation and kit thereof

Abstract

The invention discloses an ARMS (Amplification Refractory Mutation System) primer for detecting human EGFR, KRAS and BRAF gene mutation. The ARMS primer is characterized in that sequences of the ARMS primer are as shown as in SEQ ID NO.1 to SEQ ID NO.31. The invention also discloses a kit containing the ARMS primer. According to the ARMS primer disclosed by the invention, a double-internal reference system is guided in an ARMS real-time fluorescence quantification technology, a normal site reference which is consistent with a mutation site is added for contrast expect a common internal reference, the defects of the prior art are avoided by the double-internal reference system and a corresponding double interpretation rule, the detecting accuracy is increased, and the error rate is reduced; a reagent which is used for carrying out Sanger sequencing on a sample is also contained in the detecting kit; by combining a Sanger sequencing technology with the ARMS real-time fluorescence quantification technology, a mutation site of a known gene can be detected, a new mutation site can also be found, and the detecting sensitivity is ensured.

Description

technical field [0001] The invention relates to a primer for detecting human EGFR, KRAS and BRAF gene mutations and a kit thereof, belonging to the technical field of molecular biology detection. Background technique [0002] According to the recommendation of NCCN, an authoritative organization in the field of cancer research and treatment in the world, lung cancer patients, especially non-small cell lung cancer (NSCLC) patients, are recommended to undergo EGFR gene mutation testing. The presence of activating mutations in EGFR has an important role in the selection of anticancer drugs for treatment. Studies have shown that some mutations of EGFR, especially its exon 19 deletion mutation, exon 21 point mutation (L858R), exon 18 point mutation, exon 20 point mutation (T790M), and The efficacy of tyrosine kinase inhibitors (TKIs) is highly correlated. In addition, in some lung cancer patients, mutations of EGFR gene and KRAS gene exist at the same time, and mutations in exo...

AI Technical Summary

Problems solved by technology

Although the use of fluorescent probe technology can improve the specificity of detection, the cost of use is too high, and new mutations cannot be detected

In addition, the fluorescent probe is used to judge the Ct value of the fluorescence peak of the mutant group probe. Its defect is that there is no effective reference. the same gene

Moreover, the judgment rules of its Ct value are based on empirical values, so the detection effect is not ideal due to factors such as sample extraction quality during detection, and inconsistencies in detection results may occur in practical applications

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