Noninvasive biopsy virus detection method based on high throughput gene sequencing and tagged connector

A labeling and virus technology, applied in biochemical equipment and methods, microbial measurement/testing, library creation, etc., can solve the problems of incomplete reflection of the ecological composition of virus samples and unusable sequencing data, so as to reduce the cost of sequencing, High throughput and cost reduction effect

Active Publication Date: 2016-05-11
GUANGZHOU SAGENE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the conventional single nucleic acid type sequencing method used also has defects: when building a DNA library, RNA is regarded as contamination and removed, while when RNA is constructed, DNA is regarded as an impurity and removed, and the final measured DNA and ...

Method used

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  • Noninvasive biopsy virus detection method based on high throughput gene sequencing and tagged connector
  • Noninvasive biopsy virus detection method based on high throughput gene sequencing and tagged connector
  • Noninvasive biopsy virus detection method based on high throughput gene sequencing and tagged connector

Examples

Experimental program
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Embodiment 1

[0078] The material used in this example is a mixture of 8 respiratory adenovirus and syncytial virus (RSV) samples provided by Zhujiang Hospital of Southern Medical University (simulating respiratory nasopharyngeal secretions with virus infection, and the nucleic acid types are double-stranded DNA and single-stranded DNA respectively). stranded RNA). 8 mixtures were used as 8 groups of samples, each group of samples had 10 repetitions, and a total of 80 reactions were performed. In library construction, each sample was added with a DNA tag adapter. That is, add Barcode01-10 to the first group of samples; add Barcode11-20 to the second group of samples; add Barcode21-30 to the third group of samples, and so on to Barcode80. In this example, 80 DNABarcode tag adapters were combined and verified. The present invention is not limited to these 80 tag adapter sequences, and any tag adapter sequence or any combination of multiple tag adapter sequences falls within the scope of the ...

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Abstract

The invention provides a noninvasive biopsy virus detection method based on high throughput gene sequencing. The method can carry out DNA sequencing and RNA sequence in parallel and thus precisely determines the biological composition and structure of viruses in patients. The method can maximally avoid the interference of nucleic acid of host cells, effectively reduce the cost of sequencing, and is capable of detecting multiple unknown crossed viruses at the same time. 80 DNA tagged connectors are designed, related reagents are assembled to form a library so as to construct a kit, and if the throughput allows, the kit can perform sequencing on 80 samples in parallel. The experiment design is strict, the automation degree is high, the virus identification can be finished in batches within 24 hours, the requirements of scientific research and clinical detection can be met in different levels, and references are provided for clinical diagnosis and prescription of diseases related with virus infection.

Description

technical field [0001] The invention relates to a virus detection method, in particular to a method for non-invasive biopsy virus based on high-throughput gene sequencing and the used label linker. Background technique [0002] Viruses are mainly divided into DNA viruses, RNA viruses and protein viruses, which exist in almost all organisms in a heterotrophic parasitic manner, and some viruses can cause harm or even death to the host. In various environments, ecosystems are often composed of a variety of viruses, cross-infection and cause concurrent diseases, and the nucleic acid genetic material of these viruses can exist in four types: double-stranded DNA, single-stranded DNA, double-stranded RNA and single-stranded RNA . [0003] Traditional virus identification methods include filter screening, tissue culture, electron microscope observation, serology, vaccination, and cell culture observation. However, these methods all rely on information such as the isolation and enr...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/10C40B50/06C12Q1/70C12Q1/68
CPCC12Q1/6806C12Q1/6869C12Q2525/191C12Q2535/122C12Q2563/179C12Q2531/113
Inventor 伍泳彰曾宏彬陈杰
Owner GUANGZHOU SAGENE BIOTECH
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