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Device for detecting somatic cell mutation by circulating tumor DNA (deoxyribonucleic acid) samples

A technology for somatic cell mutation and sample detection, which is applied in the field of devices for detecting somatic cell mutation, and can solve problems such as micro-free DNA content.

Pending Publication Date: 2017-06-13
ANNOROAD GENE TECH BEIJING +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, the content of cell-free DNA in plasma is very small, fragmentation is serious, and circulating tumor DNA only accounts for 0.02% to 50% of the total amount of cell-free DNA in plasma. How to distinguish real SNV from PCR errors and false positives in next-generation sequencing and the noise caused by inaccurate comparisons are currently a major problem

Method used

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  • Device for detecting somatic cell mutation by circulating tumor DNA (deoxyribonucleic acid) samples
  • Device for detecting somatic cell mutation by circulating tumor DNA (deoxyribonucleic acid) samples
  • Device for detecting somatic cell mutation by circulating tumor DNA (deoxyribonucleic acid) samples

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Embodiment 1

[0094] Example 1 The device of the present invention for detecting somatic mutations using circulating tumor DNA samples

[0095] The device for detecting somatic mutations using circulating tumor DNA samples in Example 1 has:

[0096] The data acquisition module is used to acquire the DNA sequencing data of the circulating tumor DNA sample and the DNA sequencing data of the healthy population, the sequencing data includes the mutation frequency of each site in the DNA of the circulating tumor DNA sample, and the DNA sequencing data of the circulating tumor DNA sample. Each site corresponds to the mutation frequency of each individual site in the DNA of each individual in the healthy population; usually, the DNA sequencing data of the circulating tumor DNA sample comes from the data obtained by sequencing the DNA of the circulating tumor DNA sample to be tested, and the healthy population The DNA sequencing data comes from the established healthy population DNA database;

[0...

Embodiment 2

[0126] Somatic mutation detection in a peripheral blood sample from a male patient with non-small cell lung cancer.

[0127] 1.1 Extraction of cfDNA from peripheral blood samples

[0128] Blood cfDNA was extracted using MagMAX Cell-Free DNA Isolation Kit Kit (Life Company), and the extracted cfDNA was obtained. The extraction method was referred to the manual.

[0129] 1.2 End Repair

[0130] (1) Take out the required reagents from the kit stored at -20°C in advance, and see Table 1 for the preparation amount of a single sample.

[0131] Table 1

[0132]

[0133] (2) End repair reaction: After adding the DNA sample, place the 1.5mL centrifuge tube in a Thermomixer for 30 minutes at 20°C. After the reaction, use 1.8×nucleic acid purification magnetic beads to recover and purify the DNA in the reaction system, and dissolve it in 32 μL EB.

[0134] 1.3 Add "A" at the end (A-Tailing)

[0135] (1) Take out the required reagents from the kit stored at -20°C in advance, and s...

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Abstract

The invention relates to a device for detecting somatic cell mutation by circulating tumor DNA (deoxyribonucleic acid) samples. The device comprises a data acquisition module, a mutation frequency statistics module, a comparison module, a judgment module and a detection result output module. The device for detecting the somatic cell mutation by the aid of the circulating tumor DNA samples has the advantages that system errors can be accurately differentiated from the real somatic cell mutation by the device, accordingly, the sensitivity can be improved, and false positive and false negative can be reduced.

Description

technical field [0001] The invention relates to the field of low-frequency mutation detection, in particular to a device and method for detecting somatic cell mutations using circulating tumor DNA samples. Background technique [0002] Tumor cells will release genomic DNA into the blood, and these mutated DNA will also be released into the peripheral blood, which is called circulating tumor DNA (ctDNA). Genomic DNA mutation of tumor cells is a type of somatic mutation (SNV). It has been reported in the literature that the concentration of free DNA fragments with a size of 100 to 400 bp in the plasma of cancerous people is significantly higher than that of normal people, which can be used as a marker for screening. Another study found that circulating tumor DNA has already begun to appear in the blood of patients with early-stage carcinoma in situ (primary cancer). Due to the short half-life of circulating DNA in peripheral blood, circulating tumor DNA can truly reflect the...

Claims

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Application Information

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IPC IPC(8): G06F19/22
CPCG16B30/00
Inventor 陈玥茏侯光远蔡丽丽李雪峰方真玄兆伶李大为梁峻彬陈重建
Owner ANNOROAD GENE TECH BEIJING
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