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Liver cancer detection panel based on next-generation sequencing technology, kit and application of panel

A second-generation sequencing technology and detection kit technology, applied in the field of liver cancer detection panels and kits, can solve problems such as differences and achieve the effect of reducing costs

Pending Publication Date: 2021-04-09
FUDAN UNIV SHANGHAI CANCER CENT
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the heterogeneity of tumor cells, the mutations of tumor cells in a tumor tissue are also different, which has become a key problem in realizing precision medicine by finding mutant genes to use targeted drugs

Method used

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  • Liver cancer detection panel based on next-generation sequencing technology, kit and application of panel
  • Liver cancer detection panel based on next-generation sequencing technology, kit and application of panel
  • Liver cancer detection panel based on next-generation sequencing technology, kit and application of panel

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Experimental program
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Effect test

Embodiment

[0031] Step 1. Take a patient's cancer tissue and blood samples to extract DNA. The kits used are TIANGENTGuide Cells / Tissue Genomic DNA Kit and TIANGENT TGuide Large Volume BloodGenomic DNA Kit (1-3ml).

[0032] Step 2, DNA sample library preparation. First use the Bioruptor UCD-300 non-contact automatic ultrasonic breaker to randomly interrupt the DNA, ctDNA does not need this step. Take 400ng of DNA, dilute it to 50μl with nuclease-free water, transfer it to a 0.5mL Eppendorf LoBind Tube, mix well, centrifuge briefly and put it on ice for use. Place the sample: Place the centrifuge tubes symmetrically (if there is a single tube, add water to the empty tube to balance), tighten the rotor, place it on the ice box, and pre-cool for 1-2 minutes. 150-200bp ultrasonic break. Repeat the previous step of ultrasonic crushing, a total of 9 cycles, and end in about 90 minutes. The library was then prepared using the KAPA library construction kit.

[0033] Step 3, RNA probe prepara...

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Abstract

The invention belongs to the technical field of liver cancer polygene detection, and discloses a liver cancer detection panel based on a next-generation sequencing technology. Important exon regions and partial intron regions of 557 genes are enriched by using a gene probe hybridization method, and high-depth sequencing is carried out, so that events such as gene mutation and copy number variation which have clear clinical correlation with liver cancer are accurately detected; and a polygene screening list is made into a probe, and high-risk genes are detected in a targeted mode through DNA sequencing, so that gene mutation with guiding significance for diagnosis and treatment can be detected more efficiently, and important guiding significance is achieved for clinical diagnosis and discovery of targets of targeted therapy.

Description

technical field [0001] The invention belongs to the technical field of multi-gene detection of liver cancer, and relates to a liver cancer detection panel, a kit and an application thereof based on next-generation sequencing technology. Background technique [0002] Cancer is the leading cause of death and disability in the world today. Each tumor includes both genetic (germline) and tumor-specific (somatic) variations. In the past decade, high-throughput sequencing and bioinformatics technology have made great progress, and mutation information of a large number of germline and somatic genes has been collected. A large number of clinical practices have proved that the treatment of specific proteins that are mutated in cancer patients is effective. In order to achieve the goal of precision medicine, which is to give the right drug to the right patient, many laboratories have begun to apply next-generation sequencing technology in cancer genetic testing. [0003] Liver can...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/156C12Q2600/118
Inventor 王鲁赵一鸣朱卫平毛岸荣王龙蓉邵志敏黄薇胡欣施景绣裴雨晨王云锦张国亮牛珍珍
Owner FUDAN UNIV SHANGHAI CANCER CENT
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