Abstract
Severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2, a new member of the genus Betacoronavirus, is a pandemic virus, which has caused numerous fatalities, particularly in the elderly and persons with underlying morbidities. At present, there are no approved vaccines nor antiviral therapies available. The detection and quantification of SARS-CoV-2-neutralizing antibodies plays a crucial role in the assessment of the immune status of convalescent COVID-19 patients, evaluation of recombinant therapeutic antibodies, and the evaluation of novel vaccines. To detect SARS-CoV-2-neutralizing antibodies, classically, a virus-neutralization test has to be performed at biosafety level 3, considerably limiting the general use of this test. In the present work, a biosafety level 1 pseudotype virus assay based on a propagation-incompetent vesicular stomatitis virus (VSV) has been used to determine the neutralizing antibody titers in convalescent COVID-19 patients. The neutralization titers in serum of two independently analyzed patient cohorts were available within 18 h and correlated well with those obtained with a classical SARS-CoV-2 neutralization test (Pearson correlation coefficients of r = 0.929 and r = 0.939, respectively). Most convalescent COVID-19 patients had only low titers of neutralizing antibodies (ND50 < 320). The sera of convalescent COVID-19 patients also neutralized pseudotype virus displaying the SARS-CoV-1 spike protein on their surface, which is homologous to the SARS-CoV-2 spike protein. In summary, we report a robust virus-neutralization assay, which can be used at low biosafety level 1 to rapidly quantify SARS-CoV-2-neutralizing antibodies in convalescent COVID-19 patients and vaccinated individuals.
Highlights
Severe acute respiratory syndrome coronavirus type 2
The infected cells, which were maintained in the presence of Vesicular stomatitis virus (VSV) G neutralizing antibody, produced infectious pseudotype particles that harbored the corresponding spike protein in the viral envelope
Vero E6 cells by pseudotype virus was inhibited by spike-specific neutralizing antibodies and was quantified taking advantage of the firefly luciferase (FLuc) reporter protein
Summary
Severe acute respiratory syndrome coronavirus type 2 SARS-CoV-2 is readily transmitted via aerosols or droplets. It infects cells expressing the human angiotensin-converting enzyme 2 (ACE2), which serves as a cellular receptor and is essential for viral entry [4,5]. Individuals of any age may be infected with SARS-CoV-2, but the elderly and persons with underlying morbidities suffer from the disease (COVID-19), which is characterized by fever, coughing, and respiratory distress [2]. The airborne transmission, as well as the fact that humans do not possess pre-existing immunity to this virus, allowed SARS-CoV-2 to spread rapidly in the human population, causing a still ongoing pandemic
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