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CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) system for visually detecting children influenza virus and method and application thereof

A kind of influenza virus and virus technology, applied in the field of CRISPR system for visual detection of children's influenza virus, can solve the problems of not realizing rapid and simple detection, achieve the effect of reducing severe morbidity and mortality, simple operation and improving efficiency

Pending Publication Date: 2022-04-29
SHENZHEN CHILDRENS HOSPITAL +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] CRISPR is a very popular highly sensitive and specific detection technology in recent years. There are related technologies based on CRISPR for nucleic acid detection. Although the use of CRISPR technology solves the problem of high sensitivity and specific detection, after the detection is completed, it is necessary to rely on ultraviolet light. light, gel imager, or fluorescence microscope for visual interpretation, but still has not achieved the purpose of rapid and easy detection

Method used

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  • CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) system for visually detecting children influenza virus and method and application thereof
  • CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) system for visually detecting children influenza virus and method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0069] In this example, a CRISPR system for the visual detection of influenza virus in children is designed, refer to figure 1 to describe.

[0070] see figure 1 , the CRISPR system for visual detection of influenza virus in children includes virus collection system, nucleic acid amplification system and detection system;

[0071] The virus collection system includes a flow cell 111 and a virus collection element 112, the nucleic acid amplification system includes a virus lysis element 120 and a constant temperature recombinase polymerase amplification element 121, and the detection system includes a CRISPR reaction element 130 and a lateral flow test strip 131;

[0072] The flow cell 111 is connected with the virus collection element 112; the virus collection element 112 is communicated with the virus lysis element 120; the virus lysis element 120 is connected with the constant temperature recombinase polymerase amplification element 121, and is separated by the first polyvi...

Embodiment 2

[0075] In this example, four viruses (H1N1, H2N2, influenza virus B and novel coronavirus) were detected using the CRISPR system for visually detecting children's influenza virus in Example 1. The specific process is as follows:

[0076] (1) Preparation of virus lysis reaction system: 5mM Tris(hydroxymethyl)aminomethane-hydrochloric acid (Tris-HCl, pH7.5), 1% Triton X-100 (Triton-X100), 1% ethylbenzene Polyethylene glycol (NP-40), 0.2% 3-[3-(cholamidopropyl)dimethylamino]propanesulfonic acid inner salt (CHAPS), 100g / mL lysozyme and 5% sucrose The virus lysate is mixed evenly; after the above lysate is placed in a freeze dryer for 4 hours and freeze-dried, a solid-phase reaction system is obtained, and inserted into the virus lysing element 120;

[0077] (2) Prepare nucleic acid amplification reaction system: use E. coli prokaryotic expression system to express DNA polymerase, primer-binding recombinase and single-stranded DNA binding protein required in the constant temperatur...

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Abstract

The invention belongs to the field of nucleic acid detection, and discloses a CRISPR (clustered regularly interspaced short palindromic repeats) system for visually detecting children influenza virus as well as a method and application of the CRISPR system. According to the CRISPR system, a virus collection system, a constant-temperature nucleic acid amplification system and a detection system are integrated together, the detection system integrates a CRISPR reaction element and a lateral chromatography test strip, collected viruses are sequentially subjected to the steps of virus lysis, nucleic acid amplification and detection, and a detection result is visually presented on the lateral chromatography test strip through strip color development. The detection time is short, the operation is convenient, the high-sensitivity, specific and multi-channel visual detection on the children influenza virus is realized, and meanwhile, the rapid on-site detection is also facilitated.

Description

technical field [0001] The invention belongs to the field of nucleic acid detection, and in particular relates to a CRISPR system for visually detecting children's influenza virus and its method and application. Background technique [0002] Influenza (abbreviated as influenza) is one of the major public health problems faced by human beings. It is an acute respiratory infectious disease caused by influenza virus. Its epidemic characteristics are sudden outbreak, rapid spread, and wide spread. Children are a high-risk group of influenza and a high-risk group for severe cases. This infectious disease has a sudden onset and lacks specific clinical manifestations. Novel coronavirus pneumonia is an acute respiratory infectious disease caused by a new type of coronavirus infection. It has many similarities with the symptoms of influenza, and it is difficult to quickly and accurately distinguish it. The rapid and accurate detection of pathogen markers of related diseases is influe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6804C12Q1/6844C12Q1/70C12R1/93
CPCC12Q1/6804C12Q1/6844C12Q1/701C12Q2531/119C12Q2521/507C12Q2521/327C12Q2525/161C12Q2565/625C12Q2563/131
Inventor 李德发谢中建陈挚张晗裴悦李腾
Owner SHENZHEN CHILDRENS HOSPITAL
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