Method for dual-end correction of circulating tumor DNA sequencing data

A DNA sequencing and sequencing data technology, applied in biochemical equipment and methods, sequence analysis, and microbial determination/inspection, etc., can solve problems such as inability to accurately detect cancer gene mutations, improve application value and reduce false positive rates. , the effect of improving the accuracy

Pending Publication Date: 2020-06-23
HANGZHOU HEYI GENE TECH
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AI Technical Summary

Problems solved by technology

Due to the heterogeneity of tumors and complex gene mutations, traditional detection methods cannot accurately detect cancer-related gene mutations

Method used

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  • Method for dual-end correction of circulating tumor DNA sequencing data
  • Method for dual-end correction of circulating tumor DNA sequencing data

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0040] The technical scheme of the present invention will be further described below by specific examples:

[0041] (1) cfDNA extraction, targeted capture library construction and sequencing:

[0042] Using the magnetic bead method to extract cfDNA in the sample plasma for sample library construction; the sample plasma is derived from human plasma;

[0043] Sequencing adapters are added to both ends of the cfDNA molecule, and the sequencing adapters contain 8bp tag sequences for off-machine data splitting, using liquid-phase molecular probes for hybridization capture, capturing target DNA fragments, and completing library construction;

[0044] The constructed library was sequenced using a high-throughput sequencer with a read length of 150 bp and double-end sequencing. The high-throughput sequencer was Illumina NextSeq CN500 sequencer, BGISEQ-100 sequencer, BGISEQ-1000 sequencer or DA8600 sequencer instrument;

[0045] (2) Quality control of sequencing data:

[0046] Split...

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Abstract

The invention discloses a method for dual-end correction of circulating tumor DNA sequencing data. The method comprises the steps of performing cfDNA extraction, performing targetted catching for establishing a gene bank, and performing sequencing; performing quality control on sequencing data; and performing dual-end correction on the sequencing data. A dual-end sequencing method is adopted, andtwo ends of the same DNA fragment are sequenced from the positive and negative directions. The ctDNA high-flux sequencing data is subjected to sequence overlap region alkali correction, because the dual-end sequencing overlap region sequence has concurrent characteristics, the sequencing error rate can be reduced, the ctDNA gene mutation detecting and analysing accuracy can be effectively improved, particularly low-enrichment gene mutation detection accuracy is improved, the false positive rate is reduced, and the application value of ctDNA detection to clinical treatment can be increased.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for double-end correction of circulating tumor DNA sequencing data. Background technique [0002] In recent years, the morbidity and mortality of tumors have continued to increase, and they are becoming younger, which has become one of the important factors that seriously threaten life and health and cause high social burden. The 5-year survival rate of cancer patients in China is about 40%, which is far behind 60% in developed countries. It can be seen that the situation of cancer prevention and treatment in China is very severe, and effective methods are urgently needed to improve the efficiency of cancer prevention and control, diagnosis and treatment, and improve the survival rate of patients. . [0003] Previous studies have shown that gene mutations play an important role in the regulation of tumor cell growth and progression. Due to the heterogeneity of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6869C12Q1/6886G16B20/20G16B20/50G16B30/10
CPCC12Q1/6869C12Q1/6886G16B20/20G16B20/50G16B30/10C12Q2600/156C12Q2523/308C12Q2525/191C12Q2535/122
Inventor 王军一肖雯叶可勇闫楠刘杰
Owner HANGZHOU HEYI GENE TECH
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