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Tumor screening kit based on high-throughput sequencing technology for free DNA in peripheral blood plasma and system and method of tumor screening kit

A tumor screening, high-throughput technology, applied in the fields of biochemical equipment and methods, sequence analysis, and microbial determination/inspection, to achieve the effect of sensitivity and high sensitivity

Active Publication Date: 2019-09-24
广州市雄基生物信息技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing studies have shown that liquid biopsy has the potential for early screening of tumors, but the curre

Method used

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  • Tumor screening kit based on high-throughput sequencing technology for free DNA in peripheral blood plasma and system and method of tumor screening kit
  • Tumor screening kit based on high-throughput sequencing technology for free DNA in peripheral blood plasma and system and method of tumor screening kit
  • Tumor screening kit based on high-throughput sequencing technology for free DNA in peripheral blood plasma and system and method of tumor screening kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 The tumor screening kit based on the high-throughput sequencing technology of peripheral blood plasma cell-free DNA provided by the present invention includes plasma cell-free DNA extraction reagent (derived from MagaBio company), library construction reagent (using Ion PlusFragmentLibrary Kit and Ion Ampliseq TM Primer Pool reagents, from Life Technologies), sequencing reagents (Ion PI TM Hi-Q TM OT2 200Kit, Ion PI TM Hi-Q TM Sequencing Kit (sourced from Life Technologies), sequencing chip (sourced from Life Technologies), description of bioinformatics analysis methods.

Embodiment 2

[0044] Example 2 A tumor early screening system based on high-throughput sequencing technology of cell-free DNA in peripheral blood, the system includes:

[0045] (1) Nucleosome information module: the nucleosome information module is used to annotate all human protein-coding gene information in UCSC's RefSeq database, and obtain the transcription start sites (TSSs) and transcription termination sites ( Genomic position of the upstream and downstream 1Kb region of TTSs) to construct a nucleosome positioning standard database;

[0046] (2) Sequencing module: the sequencing module is used for high-throughput sequencing of the free DNA in the peripheral blood plasma of the examiner, and obtains the position information of each examiner's sequence on the genome;

[0047] (3) Quality control module: The quality control module is used to remove PCR repeat sequences caused by library construction and high-throughput sequencing, remove low-quality DNA fragment sequences, and remove un...

experiment example 1

[0050] Experimental Example 1: High-throughput sequencing and result analysis of peripheral blood plasma cell-free DNA in lung cancer patients and healthy controls

[0051] Based on the peripheral blood plasma of 38 healthy people and 11 lung cancer patients, high-throughput sequencing was carried out using the kit of the present invention, and the sequencing data was analyzed for the coverage difference of the TSSs region, and it was found that there were differences in the TSSs region of lung cancer and the control group of healthy people 557 (see Table 2). Among them, the coverage of 302 regions with differential TSSs was up-regulated, and the coverage of 255 regions with differential TSSs was down-regulated. By using differentially expressed genes for unsupervised hierarchical clustering analysis, it was found that based on 557 differentially expressed genes, the samples could be clustered into two categories: lung cancer and healthy individuals, and the patterns of differ...

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Abstract

The invention discloses a tumor screening kit based on high-throughput sequencing technology for free DNA in peripheral blood plasma and a system and method of the tumor screening kit, and aims to provide a kit for early screening of single or multiple tumors with high sensitivity, non-mutation sites and specific gene dependence by using the peripheral blood. The kit comprises a plasma free DNA extraction reagent, a library construction reagent, a sequencing reagent, a sequencing chip and a biological information analysis method. The location information and differences of nucleosome footprints can be obtained in the whole genome range of the free DNA through the high-throughput sequencing of the free DNA in the peripheral blood plasma of healthy people and tumor patients and through bioinformatics analysis of the sequencing data, and then the clustering analysis of the tumor patients and the healthy control population is realized according to the differences of the nuclosome footprints, so as to distinguish tumor patients and the healthy control population. The purpose of the above screening is to obtain information on intermediate results. The invention belongs to the field of biotechnology.

Description

technical field [0001] The invention discloses a tumor screening kit, specifically, a tumor screening kit based on the high-throughput sequencing technology of free DNA in peripheral blood plasma. Quantitative sequencing technology for tumor screening kits. Background technique [0002] World Health Organization cancer research data show that in 2018, there were an estimated 18 million new cancer cases and 9.6 million cancer deaths worldwide. At the same time, the World Health Organization proposes that as long as early detection, 90% of cancers can be completely cured. However, the first diagnosis of cancer patients in hospitals all over our country only accounts for less than 10%, and more than 90% have lost the precious opportunity to obtain good curative effect. Early cancer generally has no obvious symptoms, and when the body feels obvious discomfort and detects it, it is usually in the middle and late stages of cancer, so early cancer screening is very important. ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6869G16B30/10
CPCC12Q1/6869G16B30/10C12Q2535/122C12Q2531/113
Inventor 胥顺李坤杨学习
Owner 广州市雄基生物信息技术有限公司
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