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Method for detecting coronavirus by adopting angiotensin converting enzyme II (ACE2)

An angiotensin, coronavirus technology, applied in the field of coronavirus detection

Inactive Publication Date: 2021-07-23
ACCUCISE DIAGNOSTICS INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Comparing the long-term practice of infectious diseases caused by pathogens and the current response to the COVID-19 pandemic, an obvious technical shortage is the detection of viral antigen (antigen) substances (such as S protein and its fragments) and virions or Detection of the virion itself

Method used

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  • Method for detecting coronavirus by adopting angiotensin converting enzyme II (ACE2)
  • Method for detecting coronavirus by adopting angiotensin converting enzyme II (ACE2)
  • Method for detecting coronavirus by adopting angiotensin converting enzyme II (ACE2)

Examples

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Embodiment Construction

[0035] 1. Label recombinant human ACE2 (rhACE2) with an electrically neutral ruthenium complex (NRC)

[0036] U.S. Patent US10203333 and Chinese Patent ZL 201480045420.X disclose several markers that help reduce non-specific adsorption and exhibit stronger luminescent signals under electrochemiluminescent conditions. In this example, one of the electrically neutral ruthenium complexes is selected. The compound, Ru(2,2'-bipyridine)(bathophenanthrolinedisulfonate) [4-(2,2'-bipyridin-4-yl)butanoic acid] (NRC16 for short), is used as a luminescent marker.

[0037]

[0038] 100 μg (1.1 nmol) of rhACE2 was dissolved in 500 μL of 0.01 M PBS (pH 7.4) buffer. Using EDC (1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride) and sulfo-NHS (N-hydroxysulfosuccinimide sodium salt) in 2-(N-morpholine)ethanesulfonic acid buffer ( MES buffer) to activate the carboxyl group on NRC16. According to the set labeling reaction ratio, that is, the reaction concentration molar ratio of rh...

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Abstract

The present invention relates to a method for the detection or quantitative analysis of coronavirus particles and its spike protein or any RBD-containing spike protein fragment using ligand-receptor interaction, i.e. The strong binding effect of the receptor binding domain (RBD) of the coronavirus spike protein (S protein) and angiotensin converting enzyme II (ACE2). As a necessary alternative to antibodies in conventional immunoassay methods, ACE2 can be used to coat the solid phase surface to capture or enrich the test substance, and can also be coupled to a marker involved in signal generation. Depending on the property and the signal mode of the detected object, the ACE2 can be independently used as a recognized body of the detected object and can also be used in combination with an antibody.

Description

technical field [0001] The invention belongs to the field of coronavirus detection methods. Background technique [0002] The envelope diameter of SARS-CoV-2 is about 85 nanometers, and its crown spike protein (Spike protein, referred to as S protein, see figure 1 ) about 20 nm long. A large number of studies have proved that SARS-CoV-2, like the pathogen SARS-CoV of SARS, binds to the same receptor-angiotensin-converting enzyme II (ACE2) through the S protein, and binds to the transmembrane serine protease (TMPRSS2) Assisted, while invading human cells. The S protein is a trimer consisting of two subunits, S1 and S2. The S1 subunit forms the top of the crown spike, while the S2 subunit is anchored to the viral envelope. The receptor binding domain RBD on the S1 subunit (receptorbinding domain, RBD, see figure 1 Right) After binding to the human cell surface receptor ACE2, due to the action of the protease TMPRSS2, the specific site of the S protein is cut, resulting in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573G01N33/569
CPCG01N33/573G01N33/56983G01N2333/8103G01N2333/165
Inventor 周明
Owner ACCUCISE DIAGNOSTICS INC