ELIPSE-COL: A novel ELISA test based on rational envisioned synthetic peptides for detection of SARS-CoV-2 infection in Colombia

  1. Adriana Arévalo
  2. Carlos Franco-Muñoz
  3. Sofía Duque-Beltrán
  4. Lyda Muñoz-Galindo
  5. María Herrera-Sepulveda
  6. José Manuel Lozano
  7. Luz Mary Salazar
  8. Martha L. Ospina-Martinez
  9. Marcela Mercado-Reyes

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Abstract

Background

COVID-19 pandemic caused by infection with the betacoronavirus SARS-CoV-2 is the greatest public health defiant on a global scale in the last 100 years. Governments and health Institutes face challenges during the pandemic, related to the diagnosis, mitigation, treatment, and timely detection after the epidemic peak for the prevention of new infections and the evaluation of the real impact of the COVID-19 disease in different geographic areas. To develop a valuable tool to study the seroprevalence of SARS-CoV-2 infection in Colombia, an “ in-house ” ELISA was achieved for the detection of IgG anti-SARS-CoV-2 antibodies in serum.

Methods

The test was standardized using an antigenic epitope “Pool” of the synthetic peptide as antigen derived from antigenic regions of the spike, nucleocapsid, envelope, and membrane structural proteins, which were designed, based on the genomic information of SARS-CoV-2 circulating in Colombia. In the ELISA standardization process, 94 positive sera were used, including sera from asymptomatic and symptomatic patients (mild and severe) and 123 negative sera, including pre-pandemic historical negatives originating from patients living in arbovirus endemic areas or patients with a history of respiratory diseases and sera from patients with a negative rRT-PCR test for SARS-CoV-2.

Results

The in-house peptide ELIPSE-COL test showed promising performance, being able to detect reactivity in sera from asymptomatic and symptomatic patients. The sensitivity and specificity of the assay were 91.4% and 83.7% respectively.

Conclusion

ELIPSE-COL assay was developed as an ELISA test using synthetic peptides for the study of the seroprevalence of SARS-CoV-2 infection in Colombia.

SUMMARY BOX

  • Detection of IgG anti-SARS-CoV-2 antibodies is required for the evaluation of the pandemic impact and vaccination strategies.

  • ELIPSE-COL is an in-house test based on synthetic peptides as antigen derived from antigenic regions of the spike, nucleocapsid, envelope, and membrane structural proteins.

  • The sensitivity and specificity of the assay were 91.4% and 83.7% respectively suggesting a promising performance.

  • ELIPSE-COL test is a valuable tool for the study of seroprevalence in Colombia.

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  1. Version published to 10.1101/2020.11.13.20230060v2 on medRxiv
  2. ScreenIT

    SciScore for 10.1101/2020.11.13.20230060: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Institutional Review Board StatementIRB: Ethical considerations: This work was approved by the Ethics and Research Methodologies Committee (CEMIN) of the National Institute of Health (created by resolution 395 of April 4, 2017) who determined that the work meets the technical and ethical requirements for which they granted ethical approval registered in Act No. 7 of April 13, 2020.
    Consent: All necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    ELIPSE-COL: immunodiagnostic assay for the detection of anti-SARS-CoV-2 antibodies: Polystyrene 96-flat bottom plates (Inmulon IB) were immobilized overnight with a “Pool” of 7 proteins peptides: 2 of Spike protein peptides, 3 Nucleocapsid proteins peptides from different antigenic regions, 1 Membrane protein-peptide, and 1 Envelope protein-peptide all of them derived from antigenic regions of SARS-CoV-2.
    anti-SARS-CoV-2
    suggested: None
    Then a goat alkaline phosphatase anti-human-Ig-conjugate was poured at 1:500 of dilution on PBS-Tween-20 and incubated for one hour at 37°C to bind specific human antibodies to SARS-CoV-2 epitopes.
    alkaline phosphatase
    suggested: None
    Software and Algorithms
    SentencesResources
    Peptide Design: Worldwide and Colombian SARS-CoV-2 genome sequences were retrieved from GISAID and NCBI databases (https://www.gisaid.org/
    NCBI
    suggested: (NCBI, RRID:SCR_006472)
    Substitution matrices of nucleotides and amino acids of structural proteins were generated from a multiple sequence alignment with the SARS-CoV-2 genomes using the Muscle Algorithm (14) in MEGA X (15).
    MEGA
    suggested: (Mega BLAST, RRID:SCR_011920)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: We did not find any issues relating to colormaps.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    About SciScore

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  3. Version published to 10.1101/2020.11.13.20230060v1 on medRxiv