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Coronavirus01:29

Coronavirus

Coronaviruses, including the severe acute respiratory syndrome coronavirus (SARS-CoV), are enveloped viruses characterized by their single-stranded, positive-sense RNA genome and helical nucleocapsid structure. The hallmark of these viruses is their club-shaped spike (S) glycoproteins that protrude from the viral envelope, facilitating attachment to host cells. Typically, coronaviruses infect the upper respiratory tract, often causing mild or asymptomatic disease. However, certain strains like...

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SARS-CoV-2 nucleocapsid detection using a recombinant phage display-isolated single-chain fragment variable.

Abbasali Salarifar1, Mohammad Javad Rasaee1

  • 1Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

Journal of Immunoassay & Immunochemistry
|March 25, 2025
PubMed
Summary

Researchers developed a novel SARS-CoV-2 diagnostic tool using a single-chain fragment variable (scFv) displayed on a phage. This method overcomes immobilization challenges, offering a sensitive and specific approach for detecting the virus.

Keywords:
COVID-19SARS-CoV-2nucleocapsidphage displayrecombinant antibodies

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Area of Science:

  • Immunology
  • Molecular Biology
  • Virology

Background:

  • Accurate disease diagnosis is crucial for effective control.
  • Single-chain fragment variables (scFvs) show diagnostic potential but face immobilization limitations.
  • This study addresses scFv immobilization issues for improved diagnostic applications.

Purpose of the Study:

  • To isolate a SARS-CoV-2 nucleocapsid phosphoprotein (NP)-specific scFv.
  • To develop a diagnostic tool using scFv-displaying phage format.
  • To overcome the limitations of scFv immobilization in diagnostic assays.

Main Methods:

  • Constructed an scFv library using splicing by overlap extension (SOE) PCR from immunized mice spleen.
  • Cloned the library into a phagemid vector and screened against SARS-CoV-2 NP antigen.
  • Utilized phage display panning and ELISA to identify and assess scFv clone #31 for sensitivity and specificity.

Main Results:

  • Successfully constructed and screened an scFv-displaying phage library against SARS-CoV-2 NP.
  • Identified clone #31 as a highly qualified scFv with favorable binding capabilities.
  • Demonstrated significant sensitivity and specificity of clone #31 against the NP antigen in ELISA.

Conclusions:

  • Clone #31, an NP-specific scFv, is a promising candidate for SARS-CoV-2 diagnostics.
  • The scFv-displaying phage format effectively overcomes immobilization challenges.
  • This approach holds potential for developing advanced diagnostic tools and therapeutics for SARS-CoV-2.