Production of Sars-Cov-2 Membrane Protein as Better Target Protein for Diagnostic Application
International Journal of Infectious Diseases
; 130(Supplement 2):S139, 2023.
Article
in English
| EMBASE | ID: covidwho-2325715
ABSTRACT
Intro The COVID-19 pandemic is caused by the SARS-CoV-2 virus, an enveloped RNA of the coronavirus family. The advancement in molecular technology and biochemistry has accelerated the development of diagnostic reagents and assays. Much attention has been focused on the S protein, but the high mutation rate in this region could lead to false negative results. Thus, a better target protein for diagnostic application is needed for accurate detection. Method(s) Nucleotide sequences encoded for membrane (M) glycoprotein gene region of SARS-CoV-2 from Malaysian isolates were extracted from GISAID, aligned, and selected accordingly. The DNA plasmid was commercially synthesized with codon optimization for Escherichia coli (E. coli), and the presence of the M gene was confirmed by PCR. The plasmid was then transformed into E. coli. Later, the expression of M glycoprotein was induced, separated on an SDS-PAGE gel, and transferred onto a nitrocellulose membrane, followed by immunostaining. Finding(s) The analysis of the M glycoprotein against the Omicron strains demonstrated that the amino acid is conserved (99.5%). The M glycoprotein was successfully expressed and detected with antibodies from SARS-CoV-2 infected patients at ~26 kDa. The protein is currently upscale for the generation of monoclonal Ab (Mab). Discussion(s) The M protein of SARS-CoV-2 is more conserved among the virus and also has been reported to confer antigenic properties. Selection of M protein perhaps a better option compared to current detection assays that use spike (S) protein, which could lead to false negative results, as this gene region particularly the ribosome-binding domain (RBD) rapidly undergoes mutations. The utilization of M protein potentially improves negative predictive value (NPV) of the diagnostic test. Conclusion(s) Further development of diagnostic reagents is needed to improve the assay's specificity. The newly developed M protein and the MAb can be used to generate a more accurate viral detection assay.Copyright © 2023
adult; antigenicity; bacterial strain; codon; conference abstract; controlled study; diagnosis; diagnostic test accuracy study; Escherichia coli; false negative result; human; immunohistochemistry; nonhuman; nucleotide sequence; plasmid; polyacrylamide gel electrophoresis; polymerase chain reaction; predictive value; protein domain; ribosome; Severe acute respiratory syndrome coronavirus 2; spike; virus detection; amino acid; endogenous compound; glycoprotein; M protein; membrane protein; monoclonal antibody; pyroxylin
Full text:
Available
Collection:
Databases of international organizations
Database:
EMBASE
Type of study:
Diagnostic study
/
Prognostic study
Topics:
Variants
Language:
English
Journal:
International Journal of Infectious Diseases
Year:
2023
Document Type:
Article
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