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Detection method and detection device for ctDNA proportion based on capture sequencing

A detection method and sequencing technology, applied in the field of genetic engineering, can solve the problem of low accuracy

Active Publication Date: 2019-06-14
ZHENYUE BIOTECHNOLOGY JIANGSU CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0011] The present invention aims to provide a detection method and detection device for ctDNA proportion based on capture sequencing, so as to solve the technical problem of low accuracy in the detection of ctDNA proportion in the prior art

Method used

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  • Detection method and detection device for ctDNA proportion based on capture sequencing
  • Detection method and detection device for ctDNA proportion based on capture sequencing
  • Detection method and detection device for ctDNA proportion based on capture sequencing

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Embodiment 1

[0178] 1. Sample Preparation

[0179] Select blood cell DNA and cfDNA samples from 6 tumor patients (respectively denoted as sample A, sample B, sample C, sample D, sample E, and sample F), and perform the following operations respectively:

[0180] 1) Purification and quantification of cfDNA

[0181] The cfDNA in the sample to be tested will be subjected to 2100 quality inspection to see if there are large fragments. For the purification of cfDNA samples containing large fragments, since the magnetic beads have the characteristic of first adsorbing large fragments, first use 0.5 times magnetic beads to purify, at this time, large fragments are adsorbed on the magnetic beads, and the supernatant is sucked into 2.5 times magnetic beads, All products except large fragments were recovered. Qubit quantification was performed on all samples.

[0182] 2) Purification and quantification of blood cell DNA

[0183] Take a sufficient amount of 6 blood cell DNA in the sample to be te...

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Abstract

The invention discloses a detection method and detection device for ctDNA (circulating tumor DNA) proportion based on capture sequencing. The detection method includes the following steps: S1, obtaining baseline sample DNA and gene data of capture sequencing of cfDNA (circulating cell-free DNA) to be tested; S2, using the homozygous and heterozygous sites in the baseline sample DNA at the same time, and selecting the sites on which the mutation frequency between the baseline sample DNA and the cfDNA to be tested is significantly different and the predetermined filtering conditions are satisfied as candidate SNP (Single Nucleotide Polymorphisms) sites; S3, determining a mutation type of the candidate SNP sites of normal cell DNA and ctDNA, in combination with the situation that copy numbervariation occurs in the region where the candidate SNP sites are located; and S4, establishing a probability model by using a maximum likelihood method, and obtaining ctDNA proportion by calculation through the probability model. The application of the technical scheme of the detection method and detection device for ctDNA proportion based on capture sequencing can improve the sensitivity and accuracy of CNV (copy number variation) detection of plasma ctDNA at the same time from various aspects.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a detection method and a detection device based on the proportion of ctDNA captured and sequenced. Background technique [0002] Circulating tumor DNA (circulating tumor ctDNA) is produced by tumor cell apoptosis, necrosis or secretion, and is a kind of circulating cell-free DNA (cfDNA). ctDNA has a short half-life in blood and can be used for real-time monitoring of tumor patients. In addition to ctDNA single nucleotide polymorphisms (Single Nucleotide Polymorphisms, SNP), insertion-deletion markers (insertion-deletion, InDel) and copy number variation (copynumber variation, CNV), the proportion of ctDNA in cfDNA can also be used as An indicator of tumor progression and prognosis. According to the proportion of ctDNA, the detection results of SNP, InDel and CNV can also be corrected. [0003] Like tumor tissue, ctDNA cannot be used to detect the proportion of tumo...

Claims

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Application Information

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IPC IPC(8): G16B40/00G16B20/20G16B20/50G16B20/10
Inventor 韩天澄于佳宁侯军艳林小静陈维之杜波何骥
Owner ZHENYUE BIOTECHNOLOGY JIANGSU CO LTD
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