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Virus integrated DNA enrichment method, sequencing data analyzing method and device

A sequencing data, integrated technology, applied in the field of virus integration DNA enrichment, can solve the problems of restricted application, high probe cost, insufficient specificity, etc., and achieve the effect of saving data volume and high sequencing depth.

Pending Publication Date: 2019-09-24
SHENZHEN HAPLOX BIOTECH
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Problems solved by technology

In the field of high-throughput sequencing, the most widely used library target enrichment technology is probe hybridization capture technology, but the disadvantages of high probe cost, long hybridization time, and insufficient specificity greatly restrict its application in virus integration. Applications in DNA high-throughput sequencing research
At the same time, the heterozygous fragments of viral genomic DNA and human genomic DNA are also cumbersome in the processing and analysis of sequencing data

Method used

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  • Virus integrated DNA enrichment method, sequencing data analyzing method and device
  • Virus integrated DNA enrichment method, sequencing data analyzing method and device
  • Virus integrated DNA enrichment method, sequencing data analyzing method and device

Examples

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Embodiment

[0071] In this example, specific primers, namely capture primers, are designed and artificially synthesized based on the genome sequence information of the virus; the capture primers are enriched with the next-generation sequencing pre-library to be enriched, and the hybridization products are adsorbed with affinity streptomycin magnetic beads after the reaction , and the non-specifically adsorbed nucleic acids were removed by washing with the washing solution. The magnetic beads adsorbed with hybridization products were subjected to POST-PCR, and then sequenced on the machine. After the data is off the machine, filter the original data of double-end sequencing to remove adapter sequences and unqualified bases to obtain clean data; then splicing and assembling R1 and R2 reads, and assembling and growing reads that overlap R1 and R2 Fragment reads; then construct a mixed reference genome of virus and human, and further perform BWA comparison; extract soft clip reads that can be...

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Abstract

The application discloses a virus integrated DNA enrichment method, a sequencing data analyzing method and a device. The enrichment method of the application comprises the steps of primer hybridizing and extending, magnetic bead separating and washing. The sequencing data analyzing method disclosed by the application comprises the steps of data filtering, splicing and assembling, comparing and breakpoint judging and gene noting. According to the enrichment method disclosed by the application, specific primers are adopted for extending virus genome target genes so as to replace a long probe, so that the cost is low, the efficiency is high, the specificity is high, and the enrichment time is shortened. Compared with a complete genome, the sequencing data analyzing method disclosed by the application can save data amount, can obtain higher sequencing depth, can obtain more virus DNA breakpoint information and integration position information on the host genome, and can obtain clonal integration events supported by more reads, and besides, can detect non-clonal integration supported by single reads, so as to construct a comprehensive prospect integrated on the host genome.

Description

technical field [0001] The present application relates to the technical field of virus-integrated DNA detection, in particular to a method for enriching virus-integrated DNA, a sequencing data analysis method and a device. Background technique [0002] Oncogenic DNA viruses, or DNA oncogenic viruses, can cause tumors in humans and animals. DNA tumorigenic viruses that have been identified as key factors in the occurrence and development of human malignant tumors include hepatitis B virus (hepatitis B virus, HBV), human papillomavirus (Human Papillomavirus, HPV), African lymphoma virus (Epstein-Barrvirus, EBV) and Kaposi's sarcoma-associated herpesvirus (Kaposi's sarcoma-associated herpesvirus, KSHV), etc. Oncogenic DNA viruses generally have a means of causing cancerous cells, that is, the integration of viral DNA on the host cell genome to form Novel viral integrated DNA. [0003] The integration of viral DNA into the host cell genome can cause the instability of the host...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6806G16B30/10G16B20/00C12R1/93
CPCC12Q1/6806C12Q1/705C12Q1/706C12Q1/708G16B20/00G16B30/10C12Q2565/537C12Q2533/101C12Q2531/113C12Q2535/122
Inventor 陈实富林海久刘明
Owner SHENZHEN HAPLOX BIOTECH
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