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Diagnostic marker and application thereof in COVID-19 diagnosis and coronavirus previous infection detection

A technology of COVID-19 and COVID19-V004, applied in the field of biomedicine, can solve the problems of difficulty in protein structure expression, difficult to stably preserve, increase the difficulty of protein expression and purification, etc., and achieve the effect of easy operation.

Pending Publication Date: 2021-11-19
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, when preparing key binding domains such as S protein or S1-RBD, it is usually the most difficult step to express the protein with the correct structure, and viral proteins often have multiple glycosylation sites, which increases the efficiency of protein expression and purification. Difficulty (Lancet2020Apr 4;395(10230):1101-1102)
Therefore, the preparation cost is high and it is not easy to store stably

Method used

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  • Diagnostic marker and application thereof in COVID-19 diagnosis and coronavirus previous infection detection
  • Diagnostic marker and application thereof in COVID-19 diagnosis and coronavirus previous infection detection
  • Diagnostic marker and application thereof in COVID-19 diagnosis and coronavirus previous infection detection

Examples

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Embodiment 1

[0068] Example 1 Detection of serum serum with COVID-19 rehabilitation patients using a small peptide chip

[0069] 1. Processing and coupling of peptides

[0070] 1.1 Preparation: According to 12AA per peptide, there is a total of 6AA overlapping lengths, and the S protein has a total of 220 peptides. The S1 region contains 118 peptides, and the S2 region includes 102 peptides, including the peptide nraltgiaveqd and IAVEQDKNTQEV in the present invention, which is finally synthesized and purified by Gil Biochemical (Shanghai) Co., Ltd., and the N-terminal of each peptide after purification. Cys is coupled to BSA, co-coupling successful 197 (coupling products).

[0071] The specific coupling steps are as follows:

[0072] 1 Take 10 mg of BSA is dissolved in 1 mlpbs buffer, the concentration is 10 mg / ml.

[0073] 2 Take 10 μl of SMCC (preferably 1 mg SMCC is dissolved in 10 μLDMSO) is dissolved in a BSA solution, and 1 h is placed at 25 ° C.

[0074] 3 Transfer the activated BSA-...

Embodiment 2

[0147] 1, and coupling the peptide processing

[0148] Peptide COVID19-V004 (amino acid sequence: NRALTGIAVEQDKNTQEV) of GL Biochem (Shanghai) Co., synthesized and purified, together with the N-terminal Cys conjugated to the BSA, to obtain the coupling product. DETAILED same coupling procedure as in Example 1.

[0149] 2, ELISA validation peptide

[0150] Carried out using the method (step 2.3) the same as in Example 1.

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Abstract

The invention discloses a diagnostic marker and application thereof in COVID-19 diagnosis and coronavirus previous infection detection. The diagnostic marker comprises a peptide fragment COVID19-V004, the amino acid sequence of the peptide fragment COVID19-V004 is a sequence containing five or more continuous amino acids in NRALTGIAVEQDKNTQEV, or, the amino acid sequence of the peptide segment COVID19-V004 is a sequence formed by substitution or / and deletion or / and addition of one to several amino acids in the NRALTGIAVEQDKNTQEV. Based on the diagnostic marker provided by the invention, the level of the IgG antibody of the anti-peptide fragment in human serum is qualitatively detected by applying an indirect method. The detection kit established based on the invention can be used as an auxiliary means for diagnosis of COVID-19.

Description

Technical field [0001] The present invention belongs to the field of biotechnology medicine, in particular to a diagnostic marker and its application in diagnosis and COVID19 coronavirus infection detection in the past, particularly to a peptide and derivatives thereof COVID19-V004 diagnosis novel coronavirus application kit pneumoniae (COVID-19). Background technique [0002] Novel coronavirus World Health Organization in 2020 named January 12 as the 2019-nCoV, followed by the International Committee of the virus according to taxonomic research in gene sequencing will be named as "severe acute respiratory syndrome coronavirus 2 (SARS-CoV- 2) ", the World Health Organization disease caused by a virus named" 2019 coronavirus disease (COVID-19) ". 2020 January 30, World Health Organization announced the new coronavirus constitute a public health emergency of international concern. [0003] SARS-CoV-2 is a novel β is a new strain of coronavirus, can cross the species barrier to infe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/58G01N33/569G01N33/543
CPCG01N33/6854G01N33/581G01N33/56983G01N33/54306G01N2333/165G01N2469/20
Inventor 陶生策李阳赖丹昀江何伟张海南祁环
Owner SHANGHAI JIAO TONG UNIV