Hybrid capture kit and method for detecting tumor individually-medicated sixteen gene hot spots

Abstract

The invention discloses a hybrid capture kit for detecting tumor individually-medicated sixteen gene hot spots. The hybrid capture kit comprises a hybridization reagent, a PCR amplification reagent and an enrichment degree detection reagent, wherein the hybridization reagent comprises an SEQ NO.1 to SEQ NO.396 probe mixture, a molar ratio of every two probes is 1:1, and the use volume of the probemixture with the concentration being 10 nM each during detection is 1 [mu]l. The kit is gene detection based on a second-generation sequencing technology, can detect three variation types (mutation,insertion deletion and fusion) one time, and uses a high sequencing depth to cover micro-scale gene variation in order to accurately detect the mutation condition of a patient on the premise of limited samples, and the mutation frequency of 1% can be accurately detected. Polygene and high-sensitivity parallel detection improves the detection rate of medicated gene variation, so the medication opportunity is not missed; and hot spots, rare and even unknown gene variation cane be detected during one-time detection, and relevant drug-resistant mutations (such as KRAS and ALK point mutation) are detected when drug-sensitive variation is found, so the sensitivity and drug resistance information of the medicated patient is accurately reflected.

Description

technical field [0001] The invention relates to the field of gene mutation detection, in particular to a hybrid capture kit and a detection method for detecting 16 gene hotspots in individualized medicine for tumors. Background technique [0002] With the discovery of tumor driver genes and the research and application of corresponding targeted drugs, tumor therapy has embarked on the road of individualized gene-oriented therapy. Targeted therapy has high efficiency and low side effects, which can improve the quality of life of lung cancer patients to a greater extent. However, targeted therapy drugs only act on tumor cells with specific gene mutations. For tumor cells without specific gene mutations, targeted drugs are even less effective than chemotherapy drugs. Therefore, patients must undergo corresponding genetic testing before receiving targeted therapy. [0003] The DNA derived from tumor tissue carries the unique gene characteristics of tumor cells, including gene ...

AI Technical Summary

Problems solved by technology

Among them, hybrid capture is characterized by the ability to capture exome or even larger target regions, with high coverage, high uniformity, high sequencing depth and high sensitivity, and can match extremely short template fragments or DNA with special structures , but the operation process is more complicated than multiplex PCR capture technology, and sometimes needs to rely on more specialized equipment; multiplex PCR is simple and flexible in operation, the template requirement is relatively low, the requirements for instruments are minimized, and the goal can be completed within a few hours Sequence enrichment and library construction are generally only suitable for capturing relatively small target sequences below 50Kb, and cannot detect gene rearrangements such as gene fusions or inversions, and large fragment deletions

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