Journal
BIOTECHNOLOGY LETTERS
Volume 44, Issue 12, Pages 1507-1517Publisher
SPRINGER
DOI: 10.1007/s10529-022-03316-0
Keywords
SARS-CoV-2; Lateral flow assay; COVID-19; RBD; Nucleocapsid
Categories
Funding
- Science, Technology & Innovation Funding Authority (STDF)
- The Egyptian Knowledge Bank (EKB)
- Science, Technology & innovation Funding Authority (STIFA), Egypt
- Emergency Targeted Program
- COVID -19 Emergency call [43798]
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This study developed a qualitative immunochromatographic test for detecting immunoglobulins M and G against SARS-CoV-2. The test showed good specificity and moderate sensitivity, making it valuable for surveillance programs in low-resource countries and monitoring antibody levels in vaccinated individuals.
Serological assays for SARS-CoV-2 are being utilized at an exponential rate for surveillance programs. This enterprise was designed to develop and validate a qualitative immunochromatographic test, via the Lateral Flow Assay (LFA), for detection of immunoglobulins M and G (IgM and IgG) against both nucleocapsid (N) and the receptor-binding domain (RBD) of the spike protein of SARS-CoV-2. Both targeted proteins were cloned and expressed in baculovirus expression system utilizing insect cells Sf9. The recombinant RBD and N proteins were purified and conjugated with gold nanoparticles (AuNPs) to set up the coating antigens pad. Both anti-human IgG and IgM were dispensed on nitrocellulose membrane to capture human antibodies in serum samples. A home-made dispensing system was developed to draw identical test and control lines. The validity of the developed LFA was verified by testing serum samples from 103 convalescent COVID-19 patients who were PCR positive for SARS-CoV-2 along with 28 control serum samples. The developed strips showed distinctive bands for IgM and IgG of both proteins (RBD and N) in positive samples. The sensitivity of RBD-based LFA was 70.9% and 39.8% for IgG and IgM, respectively, with a specificity of 100% for both. The N-based LFA exhibited a sensitivity of 73.8% and 35.9% for IgG and IgM, respectively, while its specificity was 75% and 100% for IgG and IgM, respectively. Our developed LFA could afford a tool for surveillance programs in low-resource countries. Moreover, it might be functional for rapid and inexpensive monitoring of the anti-SARS-CoV-2 antibodies in the sera of vaccinated individuals.
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