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Pancreatic cancer detection panel based on next-generation sequencing technology, kit and application thereof

A second-generation sequencing technology and detection kit technology, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, specific-purpose bioreactor/fermenter, etc., can solve the problem of low coverage, high cost, and no cancer issues such as guidance

Pending Publication Date: 2021-05-28
FUDAN UNIV SHANGHAI CANCER CENT
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The application of gene sequencing to determine the targeted treatment plan for patients has become more and more common in pancreatic cancer. However, the existing sequencing of pancreatic cancer either focuses on specific genes or whole exome sequencing. The former has a small number of specific genes, The coverage rate is low, and it is difficult to achieve large-scale coverage; the latter has too many genes, most of which do not play a guiding role in the treatment of pancreatic cancer, and the cost is relatively high

Method used

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  • Pancreatic cancer detection panel based on next-generation sequencing technology, kit and application thereof
  • Pancreatic cancer detection panel based on next-generation sequencing technology, kit and application thereof
  • Pancreatic cancer detection panel based on next-generation sequencing technology, kit and application thereof

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Experimental program
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Embodiment

[0033] Step 1. Take a patient's cancer tissue and blood samples to extract DNA. The kits used are TIANGENTGuide Cells / Tissue Genomic DNAKit and TIANGENT TGuide Large Volume Blood Genomic DNAKit (1-3ml).

[0034] Step 2, DNA sample library preparation. First use the BioruptorUCD-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.

[0035] Step 3, RNA probe preparati...

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Abstract

The invention belongs to the technical field of pancreatic cancer multi-gene detection, and discloses a pancreatic cancer detection panel based on a next-generation sequencing technology, events such as gene mutation, copy number variation and the like having clear clinical correlation with pancreatic cancer, a multi-gene screening list is made into a probe, high-risk genes are detected in a targeted manner through DNA sequencing, gene mutation with guiding significance on diagnosis and treatment can be detected more efficiently, mutant genes can be detected in a targeted mode, and the gene detection cost of patients can also be reduced.

Description

technical field [0001] The invention belongs to the technical field of multi-gene detection of pancreatic cancer, and relates to a pancreatic cancer detection panel, a kit and applications thereof based on next-generation sequencing technology. Background technique [0002] Pancreatic cancer is a highly malignant tumor of the digestive tract that is difficult to diagnose and treat. About 90% of them are ductal adenocarcinomas originating from the ductal epithelium. The morbidity and mortality have increased significantly in recent years. The 5-year survival rate is <1%, and it is one of the malignant tumors with the worst prognosis. The early diagnosis rate of pancreatic cancer is not high, the surgical mortality rate is high, and the cure rate is very low. [0003] The application of gene sequencing to determine the targeted treatment plan for patients has become more and more common in pancreatic cancer. However, the existing sequencing of pancreatic cancer either focu...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6858C12N15/11C12M1/34C12M1/00
CPCC12Q1/6886C12Q1/6858C12Q2600/156C12Q2600/118C12Q2600/106
Inventor 虞先濬施思魏妙艳梁晨徐近王巍
Owner FUDAN UNIV SHANGHAI CANCER CENT
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