Primer, kit and method for determining lung cancer gene mutation site based on high-flux sequencing technology

Abstract

The invention discloses a primer, a kit and a method for determining a lung cancer gene mutation site based on a high-flux sequencing technology, which belongs to the field of biological molecular detection. The invention discloses a method and a kit for determining a lung cancer gene mutation site based on a high-flux sequencing technology. The method comprises the following steps: extracting tumor tissue DNA; designing a panel of a lung cancer targeting treatment molecular diagnosis associated gene; performing the PCR primer amplification; and establishing a library, and performing the high-flux sequencing. The specific primer and the kit for determining the lung cancer gene mutation site are used for detecting 316 mutation situations of 16 oncogenes, and the mutation can be the replacement, insertion and/or deletion of one or more alkaline groups. The sensitivity of the detection method and the kit of the invention can reach up to 1 percent, the detection result is definite and objective and can directly reflect the specific mutation site of the reaction associated gene and has important significance for early diagnosis or auxiliary diagnosis and screening of the cancer and theprognosis monitoring of the cancer.

Description

technical field [0001] The invention belongs to the field of biomolecular detection, and more specifically relates to a high-throughput sequencing technology-based molecular diagnostic primer, kit and diagnostic method for targeted therapy of lung cancer. Background technique [0002] According to the latest figures released by the National Cancer Center, lung cancer ranks first in the incidence and death of malignant tumors in my country, with about 733,000 new cases and 591,000 deaths each year, accounting for more than 35% of all cancer deaths, surpassing traffic Accidents, natural disasters, etc. have become the most important factors affecting the death of our country's population. Compared with the statistical data of the past 30 years, the mortality rate of lung cancer in my country has increased by 465%. According to the "2017 American Cancer Data Survey Report" released by the American Cancer Society, the incidence and mortality of lung cancer in the United States c...

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Problems solved by technology

[0010] In view of the existing problems such as difficulty in accurately determining the mutation site of lung cancer gene, poor repeatability, many false positives and false negatives, and low throughput, the present invention provides a primer and reagent for determining the mutation site of lung cancer gene based on high-throughput sequencing technology Box and method for detecting and determining 316 mutations of 16 oncogenes, the oncogenes being AKT1, ALK, ARAF, BRAF, CDKN2A, EGFR, ERBB2, FGFR3, KIT, KRAS, MAP2K1, MTOR, NRAS , PDGFRA, PIK3CA and PTEN, the mutations can be substitutions, insertions and / or deletions of one or more bases

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