Evaluation of a multi-species SARS-CoV-2 surrogate virus neutralization test

Carmen W E Embregts, Babs Verstrepen, Jan A M Langermans, Kinga P Böszörményi, Reina S Sikkema, Rory D de Vries, Donata Hoffmann, Kerstin Wernike, Lidwien A M Smit, Shan Zhao, Barry Rockx, Marion P G Koopmans, Bart L Haagmans, Thijs Kuiken, Corine H GeurtsvanKessel

Research output: Contribution to journalArticleAcademicpeer-review


Assays to measure SARS-CoV-2-specific neutralizing antibodies are important to monitor seroprevalence, to study asymptomatic infections and to reveal (intermediate) hosts. A recently developed assay, the surrogate virus-neutralization test (sVNT) is a quick and commercially available alternative to the "gold standard" virus neutralization assay using authentic virus, and does not require processing at BSL-3 level. The assay relies on the inhibition of binding of the receptor binding domain (RBD) on the spike (S) protein to human angiotensin-converting enzyme 2 (hACE2) by antibodies present in sera. As the sVNT does not require species- or isotype-specific conjugates, it can be similarly used for antibody detection in human and animal sera. In this study, we used 298 sera from PCR-confirmed COVID-19 patients and 151 sera from patients confirmed with other coronavirus or other (respiratory) infections, to evaluate the performance of the sVNT. To analyze the use of the assay in a One Health setting, we studied the presence of RBD-binding antibodies in 154 sera from nine animal species (cynomolgus and rhesus macaques, ferrets, rabbits, hamsters, cats, cattle, mink and dromedary camels). The sVNT showed a moderate to high sensitivity and a high specificity using sera from confirmed COVID-19 patients (91.3% and 100%, respectively) and animal sera (93.9% and 100%), however it lacked sensitivity to detect low titers. Significant correlations were found between the sVNT outcomes and PRNT50 and the Wantai total Ig and IgM ELISAs. While species-specific validation will be essential, our results show that the sVNT holds promise in detecting RBD-binding antibodies in multiple species.

Original languageEnglish
Article number100313
Pages (from-to)1-8
Number of pages8
JournalOne Health
Publication statusPublished - Dec 2021


  • SARS-CoV-2
  • Serology
  • Neutralizing antibodies
  • Surrogate virus neutralization test
  • Animal sera


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