Development and performance evaluation of a rapid in-house ELISA for retrospective serosurveillance of SARS-CoV-2

• Published in: PloS one Vol. 16; no. 2; p. e0246346
• Main Authors: Sil, Bijon Kumar, Jahan, Nowshin, Haq, Md Ahsanul, Oishee, Mumtarin Jannat, Ali, Tamanna, Khandker, Shahad Saif, Kobatake, Eiry, Mie, Masayasu, Khondoker, Mohib Ullah, Jamiruddin, Mohd Raeed, Adnan, Nihad
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 02-02-2021; Public Library of Science (PLoS)
• Subjects: Antibodies; Antibodies, Viral - analysis; Antigens; Asymptomatic; Biology and life sciences; Coronavirus Nucleocapsid Proteins - analysis; Coronaviruses; COVID-19; COVID-19 - diagnosis; COVID-19 - virology; Diagnostic software; Diagnostic systems; Disease transmission; Editing; Enzyme-linked immunosorbent assay; Enzyme-Linked Immunosorbent Assay - instrumentation; Enzyme-Linked Immunosorbent Assay - methods; Evaluation; FDA approval; Health care; Health risks; Humans; Immunoassay; Immunoglobulin G - analysis; Infections; Life sciences; Medical research; Medicine and health sciences; Methodology; Methods; Microbiology; Morbidity; Performance evaluation; Phosphoproteins - analysis; Polymerase chain reaction; Proteins; Public health; Research and Analysis Methods; Research facilities; Reviews; Ribonucleic acid; RNA; RNA editing; SARS-CoV-2 - isolation & purification; Science and technology; Sensitivity and Specificity; Sentinel health events; Serology; Severe acute respiratory syndrome; Severe acute respiratory syndrome coronavirus 2; Technology; Viral diseases; Bangladesh; United States > US; China; Japan
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0246346

In the ongoing pandemic situation of COVID-19, serological tests can complement the molecular diagnostic methods, and can be one of the important tools of sero-surveillance and vaccine evaluation.
To develop and evaluate a rapid SARS-CoV-2 specific ELISA for detection of anti-SARS-CoV2 IgG from patients' biological samples.
In order to develop this ELISA, three panels of samples (n = 184) have been used: panel 1 (n = 19) and panel 2 (n = 60) were collected from RT-PCR positive patients within 14 and after 14 days of onset of clinical symptoms, respectively; whereas panel 3 consisted of negative samples (n = 105) collected either from healthy donors or pre-pandemic dengue patients. As a capturing agent full-length SARS-CoV2 specific recombinant nucleocapsid was immobilized. Commercial SARS-CoV2 IgG kit based on chemiluminescent assay was used for the selection of samples and optimization of the assay. The threshold cut-off point, inter-assay and intra-assay variations were determined.
The incubation/reaction time was set at a total of 30 minutes with the sensitivity of 84% (95% confidence interval, CI, 60.4%, 96.6%) and 98% (95% CI, 91.1%, 100.0%), for panel 1 and 2, respectively; with overall 94.9% sensitivity (95% CI 87.5%, 98.6%). Moreover, the clinical specificity was 97.1% (95% CI, 91.9%, 99.4%) with no cross reaction with dengue samples. The overall positive and negative predictive values are 96.2% (95% CI 89.2%, 99.2%) and 96.2% (95% CI, 90.6% 99.0%), respectively. In-house ELISA demonstrated 100% positive and negative percent agreement with Elecsys Anti-SARS-CoV-2, with Cohen's kappa value of 1.00 (very strong agreement), while comparing 13 positive and 17 negative confirmed cases.
The assay is rapid and can be applied as one of the early and retrospective sero-monitoring tools in all over the affected areas.