Nucleic acid aptamer of SARS-CoV-2 S protein or RBD protein and application of nucleic acid aptamer

A nucleic acid aptamer, sars-cov-2 technology, applied to the nucleic acid aptamer of SARS-CoV-2 S protein or RBD protein and its application field, can solve the problems that are difficult to use in the early battlefield of epidemic prevention and control

Active Publication Date: 2020-10-30
XIAMEN UNIV
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Problems solved by technology

However, since the most commonly used recognition molecules such as antibodies usually require a certain period of experimentat

Method used

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  • Nucleic acid aptamer of SARS-CoV-2 S protein or RBD protein and application of nucleic acid aptamer
  • Nucleic acid aptamer of SARS-CoV-2 S protein or RBD protein and application of nucleic acid aptamer
  • Nucleic acid aptamer of SARS-CoV-2 S protein or RBD protein and application of nucleic acid aptamer

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Embodiment Construction

[0012] Chemically synthesize a 76-base DNA initial library. The two ends of the DNA initial library are fixed sequences, including a random sequence of 40 bases in the middle, which is 5'-ATCCAGAGTGACGCAGCA-40N-TGGACACGGTGGCTTAGT-3', the library capacity for 10 15 above. Take 5nmol DNA initial library dissolved in binding buffer (12 mmol / L PBS, pH 7.4, 150mmol / L NaCl, 5mmol / L KCl, 0.55mmol / L MgCl 2 ) for denaturation treatment: 95°C for 10 minutes, 5 minutes on ice, and then placed at 25°C for 10 minutes.

[0013] 2) The recombinant SARS-CoV-2 RBD protein expressed in the Fc region of the mouse C-terminal IgG (the spike glycoprotein receptor binding domain, namely the S protein domain, purchased from Sino Biological) was used as the screening target of the aptamer. SARS-CoV-2 S protein or RBD protein is modified on Protein A latex microspheres, and the IgG-tag epitope interacts with Protein A to form RBD-Protein A latex microspheres.

[0014] 3) The obtained pretreated init...

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Abstract

The invention discloses a nucleic acid aptamer of SARS-CoV-2 S protein or RBD protein. The nucleic acid aptamer comprises nucleotide sequences as shown in SEQ ID NO: 1 to SEQ ID NO: 8. The aptamer provided by the invention can be used for providing rapid and accurate detection of SARS-CoV-2 S protein or RBD protein; meanwhile, the aptamer and angiotensin converting enzyme (ACE2) have a competitiveeffect, and the combination of SARS-CoV-2 S protein or RBD protein and ACE2 is efficiently blocked, so that the virus infection of cells is inhibited, and the aptamer can be used as a medicine for preventing and treating SARS-CoV-2 related diseases and becomes an antiviral potential medicine. The screening requirements of various application scenes such as families and communities are met, unnecessary patient aggregation is reduced, and an existing diagnosis and treatment system is supplemented.

Description

technical field [0001] The present invention relates to the nucleic acid aptamer of SARS-CoV-2 S protein or RBD protein and application thereof. Background technique [0002] The World Health Organization has declared the outbreak of the novel coronavirus (SARS-CoV-2 or 2019 nCoV) a global pandemic. The mechanism of coronavirus infection is not fully understood, nor is there any treatment or vaccine. [0003] In terms of virus diagnosis, a number of products have been approved, such as the new coronavirus SARS-CoV-2 nucleic acid detection kit (fluorescent PCR method) and the 2019 new coronavirus nucleic acid sequencing system. This kind of detection method based on SARS-CoV-2 nucleic acid usually needs to extract viral RNA, use the primers of characteristic fragments to perform RT-PCR or fluorescent RT-PCR, and judge whether the patient is infected or not by the result of nucleic acid amplification. This diagnostic method for viral nucleic acid has multiple advantages such...

Claims

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Application Information

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IPC IPC(8): C12N15/115G01N33/569A61K31/7088A61P31/14
CPCA61P31/14C12N15/115C12N2310/16G01N33/56983G01N2333/165
Inventor 杨朝勇宋彦龄魏心语朱琳朱志
Owner XIAMEN UNIV
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