Development of Enzyme-linked Immunosorbent Assay for Serodiagnosis of COVID-19
Keywords:
COVID-19, ELISA, SARS-CoV-2, IgM, IgGAbstract
Emerging of pandemic coronavirus disease-19 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) leading to diverse respiratory pathological disorders particularly severe pneumonia worldwide. Serological tests are essential approach to support diagnosis. We developed immunological assays based on capture ELISA to detect immunoglobulin M (IgM) and immunoglobulin G (IgG) against SARS-CoV-2 nucleocapsid protein presented in sera of COVID-19 patients. Four main conditions were optimised including concentration of capture molecule, sample dilution ratio, blocking reagent and concentration of conjugated protein. Anti-SARS-CoV-2 IgM ELISA showed 41.46% sensitivity and 99% specificity while sensitivity and specificity of anti-SARS-CoV-2 IgG ELISA were 60.98% and 100%, respectively. In addition, the cut-off thresholds were established at 3.2 for IgM ELISA and 2.2 for IgG ELISA, sensitivity of IgM and IgG assays was increased to 51.28% and 89.74% based on duration of onset, respectively. It is suggested that our developed ELISA could be used to determine antibody to SARS-CoV-2.
References
Mackenzie JS, Smith DW. COVID-19: a novel zoonotic disease caused by a coronavirus from China: what we know and what we don't. Microbiol Aust 2020; 41(1): 45-50.
Li Q, Guan X, Wu P, Wang X, Zhou L, Tong Y, et al. Early transmission dynamics in Wuhan, China, of novel coronavirus–infected pneumonia. N Engl J Med 2020; 382(13): 1199-207.
Ahmad I, Rathore FA. Neurological manifestations and complications of COVID-19: a literature review. J Clin Neurosci 2020; 77: 8-12.
World Health Organization. Novel coronavirus (COVID-19). [online]. 2020; [cited 20 May 2020]; [10 screens]. Available from: URL: https://www.who.int/thailand/emergencies/novel-coronavirus-2019.
Shereen MA, Khan S, Kazmi A, Bashir N, Siddique R. COVID-19 infection: origin, transmission and characteristics of human coronaviruses. J Adv Res 2020; 24: 91-8.
Prompetchara E, Ketloy C, Palaga T. Immune responses in COVID-19 and potential vaccines: lessons learned from SARS and MERS epidemic. Asian Pac J Allergy Immunol 2020; 38(1): 1-9.
Tufan A, GÜler AA, Matucci-Cerinic M. COVID-19, immune system response, hyperinflammation and repurposing antirheumatic drugs. Turk J Med Sci 2020; 50(SI-1): 620-32.
Coperchini F, Chiovato L, Croce L, Magri F, Rotondi M. The cytokine storm in COVID-19: an overview of the involvement of the chemokine/chemokine-receptor system. Cytokine Growth Factor Rev 2020; 53: 25-32.
Molnar C, Gair J. Adaptive immune response. In: Concepts of biology - 1st Canadian edition. [online]. 2019; [cited 20 May 2020]; [25 screens]. Available from: URL: https://opentextbc.ca/biology/chapter/23-2-adaptive-immune-response.
Alberts B, Johnson A, Lewis J, Raff M, Roberts K, Walter P. Molecular biology of the cell. 4th ed. New York: Garland Science; 2002.
Zhai P, Ding Y, Wu X, Long J, Zhong Y, Li Y. The epidemiology, diagnosis and treatment of COVID-19. Int J Antimicrob Agents 2020; 55(5): 105955. (13 pages).
Carter LJ, Garner LV, Smoot JW, Li Y, Zhou Q, Saveson CJ, et al. Assay techniques and test development for COVID-19 diagnosis. ACS Cent Sci 2020; 6(5): 591-605.
Meyer B, Torriani G, Yerly S, Mazza L, Calame A, Arm-Vernez I, et al. Validation of a commercially available SARS-CoV-2 serological immunoassay. Clin Microbiol Infect. [serial online]. 2020; [cited 20 May 2020]; XX(XX): [9 screens]. Available from: URL: https://www.clinicalmicrobiologyandinfection.com/action/showPdf?pii=S1198-743X%2820%2930368-2.
Petrova G, Ferrante A, Gorski J. Cross-reactivity of T cells and its role in the immune system. Crit Rev Immunol 2012; 32(4): 349-72.
Reed GF, Lynn F, Meade BD. Use of coefficient of variation in assessing variability of quantitative assays. Clin Diagn Lab Immunal 2002; 9(6):1235-9.
Wattanaphansak S, Asawakarn T, Gebhart CJ, Deen J. Development and validation of an enzyme-linked immunosorbent assay for the diagnosis of porcine proliferative enteropathy. J Vet Diagn Invest 2008; 20(2): 170-7.
Classen DC, Morningstar JM, Shanley JD. Detection of antibody to murine cytomegalovirus by enzyme-linked immunosorbent and indirect immunofluorescence assays. J Clin Microbiol 1987; 25(4): 600-4.
Centers for Disease Control and Prevention (CDC). COVID-19 in Thailand: watch - level 1, COVID-19 risk in Thailand is low, new cases are decreasing or stable. [online]. 2020; [cited 2020 Sep 3]; [2 screens]. Available from: URL: https://wwwnc.cdc.gov/travel/notices/watch/coronavirus-thailand
