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. 2021 May 30;11(6):994.
doi: 10.3390/diagnostics11060994.

Evaluation of a Pseudovirus Neutralization Assay for SARS-CoV-2 and Correlation with Live Virus-Based Micro Neutralization Assay

Ahmed Majdi K Tolah  1 , Sayed S Sohrab  1   2 , Khaled Majdi K Tolah  3 , Ahmed M Hassan  1 , Sherif A El-Kafrawy  1   2 , Esam I Azhar  1   2
Affiliations

Evaluation of a Pseudovirus Neutralization Assay for SARS-CoV-2 and Correlation with Live Virus-Based Micro Neutralization Assay

Ahmed Majdi K Tolah et al. Diagnostics (Basel). .

Abstract

The unusual cases of pneumonia outbreak were reported from Wuhan city in late December 2019. Serological testing provides a powerful tool for the identification of prior infection and for epidemiological studies. Pseudotype virus neutralization assays are widely used for many viruses and applications in the fields of serology. The accuracy of pseudotype neutralizing assay allows for its use in low biosafety lab and provides a safe and effective alternative to the use of wild-type viruses. In this study, we evaluated the performance of this assay compared to the standard microneutralization assay as a reference. The lentiviral pseudotype particles were generated harboring the Spike gene of SARS-CoV-2. The generated pseudotype particles assay was used to evaluate the activity of neutralizing antibodies in 300 human serum samples from a COVID-19 sero-epidemiological study. Testing of these samples resulted in 55 positive samples and 245 negative samples by pseudotype viral particles assay while microneutralization assay resulted in 64 positive and 236 negative by MN assay. Compared to the MN, the pseudotyped viral particles assay showed a sensitivity of 85.94% and a specificity of 100%. Based on the data generated from this study, the pseudotype-based neutralization assay showed a reliable performance for the detection of neutralizing antibodies against SARS-CoV-2 and can be used safely and efficiently as a diagnostic tool in a biosafety level 2 laboratory.

Keywords: SARS-CoV-2; Saudi Arabia; coronavirus; micro-neutralization assay; pseudotype.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
RLU values observed at each serial dilution, virus dilution vs. RLU readout was plotted to select the needed amount of virus.
Figure 2
Figure 2
Dilution curves for the positive samples included in the study. The curves are plots of the log sample dilution vs. the percent normalized inhibition for each dilution compared to the positive control. The curves show samples with high, medium, and low titers.
Figure 3
Figure 3
Correlation of SARS-CoV-2 Ab titers as detected by PNT (x-axis) vs. MNT (y-axis).
See this image and copyright information in PMC

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