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Nanopore three-generation sequencing detection method for plasma virology

A nanopore sequencing and detection method technology, applied in the biological field, can solve the problem of human host nucleic acid pollution target nucleic acid amount and other issues

Active Publication Date: 2020-06-09
INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention applies Nanopore third-generation nano-sequencing technology to the detection of blood virus spectrum, which is a brand-new technical field, and the core technical problem is the pollution of human host nucleic acid and the large amount of target nucleic acid required for detection

Method used

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  • Nanopore three-generation sequencing detection method for plasma virology
  • Nanopore three-generation sequencing detection method for plasma virology
  • Nanopore three-generation sequencing detection method for plasma virology

Examples

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Embodiment 1

[0062] Embodiment 1, detection method

[0063] We take 24 asymptomatic adult blood collected in April 2015 at the Beijing Red Cross Blood Center as an example to illustrate the specific implementation of the present invention in sequence.

[0064] 1. Blood sample virus enrichment, nucleic acid extraction and library nucleic acid preparation

[0065] 1) Virus enrichment in plasma samples: Take 6-8ml of whole blood containing EDTA anticoagulant from healthy adults without major clinical symptoms, centrifuge at 3000g for 15min, take the supernatant as plasma, and centrifuge at 10000g for 10min to remove cell debris. Add 0.5-6ml of the supernatant to the ultracentrifuge tube, add Hank's solution to a volume of 12ml, mix carefully, balance with a balance, select a W41 rotor, centrifuge at a speed of 28800g, and centrifuge at 4°C for 2 hours. Remove the supernatant, add Hank's solution to the bottom of the tube to dissolve, and transfer to a 1.5ml centrifuge tube, add Turbo DNase d...

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Abstract

The invention provides a nanopore three-generation sequencing detection method for plasma virology. The method comprises the following steps: preparation of viral nucleic acid in blood samples and incorporation of standard nucleic acid internal reference, construction of nanopore sequencing library, real-time sequencing of nanopore and conversion of real-time fast file, bioinformatics analysis andthe like. The method not only can rapidly detect virus group of a blood sample, but also can improve coverage rate of a whole genome sequence of virus and achieve a purpose of rapidly detecting the virus of the blood sample.

Description

Technical field: [0001] The invention relates to the field of biotechnology, and relates to a method for detecting blood viruses. Background technique: [0002] Blood biosafety is a major livelihood and public security issue, and the supply security of blood resources is the fundamental guarantee for the normal operation of the national medical and health service system. There are increasingly prominent problems such as the contradiction between blood supply and demand and the risk of transfusion-transmitted diseases in my country, and the four routine blood pathogen screenings of HBV, HCV, HIV and TP can no longer meet the safety of clinical blood use. The current research on blood virus spectrum, blood virus Distribution situation There are relatively few domestic and foreign research literatures. Therefore, the study of high-precision detection methods for blood pathogens, especially blood viruses, is an urgent problem to be solved in my country, and it is also a cutting-e...

Claims

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Application Information

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IPC IPC(8): C12Q1/6869C12Q1/6806G16B30/10G16B40/00
CPCC12Q1/6806C12Q1/6869G16B30/10G16B40/00
Inventor 张婷杨帆金奇
Owner INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
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