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Published January 1, 2022 | Version v1
Journal article Open

Development of in House ELISAs to Detect Antibodies to SARS-CoV-2 in Infected and Vaccinated Humans by Using Recombinant S, S1 and RBD Proteins

Creators

  • 1. Istanbul Univ Cerrahpasa, Vet Fac, Dept Virol, TR-34098 Istanbul, Turkey
  • 2. Istanbul Univ Cerrahpasa, Cerrahpasa Fac Med, Dept Med Microbiol, TR-34098 Istanbul, Turkey
  • 3. Bezmialem Vakif Univ, Fac Pharm, Dept Pharmaceut Microbiol, TR-34098 Istanbul, Turkey
  • 4. Istanbul Univ Cerrahpasa, Cerrahpasa Fac Med, Dept Biocehm, TR-34098 Istanbul, Turkey
  • 5. Istanbul Univ Cerrahpasa, Cerrahpasa Fac Med, Dept Pulm Dis, TR-34098 Istanbul, Turkey
  • 6. Kansas State Univ, Coll Vet Med, Dept Diagnost Med & Pathobiol, Manhattan, KS 66506 USA
  • 7. Sisli Hamidiye Etfal Training & Res Hosp, Blood Ctr, TR-34098 Istanbul, Turkey
  • 8. Biruni Labs, TR-34098 Istanbul, Turkey
  • 9. Pirbright Inst, Ash Rd, Woking GU24 0NF, England

Description

(1) Background: The aim of this study was to produce in-house ELISAs which can be used to determine SARS-CoV-2-specific antibody levels directed against the spike protein (S), the S1 subunit of S and the receptor binding domain (RBD) of S in SARS-CoV-2 vaccinated and infected humans. (2) Methods: Three in-house ELISAs were developed by using recombinant proteins of SARS-CoV-2, namely the S, S1 and RBD proteins. Specificity and sensitivity evaluations of these tests were performed using sera from SARS-CoV-2-infected (n = 70) and SARS-CoV-2-vaccinated (n = 222; CoronaVac vaccine) humans in Istanbul, Turkey. The analyses for the presence of SARS-CoV-2-specific antibodies were performed using the in-house ELISAs, a commercial ELISA (Abbott) and a commercial surrogate virus neutralization test (sVNT). We also analyzed archival human sera (n = 50) collected before the emergence of COVID-19 cases in Turkey. (3) Results: The sensitivity of the in-house S, S1 and RBD ELISAs was found to be 88.44, 90.17 and 95.38%, while the specificity was 72.27, 89.08 and 89.92%, respectively, when compared to the commercial SARS-CoV-2 antibody test kit. The area under curve (AUC) values were 0.777 for the in-house S ELISA, 0.926 for the S1 ELISA, and 0.959 for the RBD ELISA. The kappa values were 0.62, 0.79 and 0.86 for the S, S1 and RBD ELISAs, respectively. (4) Conclusions: The in-house S1 and RBD ELISAs developed in this study have acceptable performance characteristics in terms of sensitivity, specificity, AUC and kappa values. In particular, the RBD ELISA seems viable to determine SARS-CoV-2-specific antibody levels, both in infected and vaccinated people, and help mitigate SARS-CoV-2 outbreaks and spread.

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