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SHR Neuro Krebs Kardio Lipid Stoffw Microb

Lichtenegger, S; Saiger, S; Hardt, M; Kulnik, S; Wagner, GE; Kleinhappl, B; Assig, K; Zauner, A; Ober, M; Kimpel, J; von, Laer, D; Zatloukal, K; Steinmetz, I.
Development of a Rapid Live SARS-CoV-2 Neutralization Assay Based on a qPCR Readout.
J Clin Microbiol. 2022; 60(7): e0037622 Doi: 10.1128/jcm.00376-22 [OPEN ACCESS]
Web of Science PubMed PUBMED Central FullText FullText_MUG

 

Führende Autor*innen der Med Uni Graz
Steinmetz Ivo
Wagner-Lichtenegger Sabine
Co-Autor*innen der Med Uni Graz
Assig Karoline
Hardt Melina
Kleinhappl Barbara
Kulnik Susanne
Ober Michelle
Saiger Sabine
Wagner-Lichtenegger Gabriel
Zatloukal Kurt
Zauner Andrea
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Abstract:
Measuring SARS-CoV-2 neutralizing antibodies after vaccination or natural infection remains a priority in the ongoing COVID-19 pandemic to determine immunity, especially against newly emerging variants. The gold standard for assessing antibody-mediated immunity against SARS-CoV-2 are cell-based live virus neutralization assays. These assays usually take several days, thereby limiting test capacities and the availability of rapid results. In this study, therefore, we developed a faster live virus assay, which detects neutralizing antibodies through the early measurement of antibody-mediated intracellular virus reduction by SARS-CoV-2 qRT-PCR. In our assay, Vero E6 cells are infected with virus isolates preincubated with patient sera and controls. After 24 h, the intracellular viral load is determined by qRT-PCR using a standard curve to calculate percent neutralization. Utilizing COVID-19 convalescent-phase sera, we show that our novel assay generates results with high sensitivity and specificity as we detected antiviral activity for all tested convalescent-phase sera, but no antiviral activity in prepandemic sera. The assay showed a strong correlation with a conventional virus neutralization assay (rS = 0.8910), a receptor-binding domain ELISA (rS = 0.8485), and a surrogate neutralization assay (rS = 0.8373), proving that quantifying intracellular viral RNA can be used to measure seroneutralization. Our assay can be adapted easily to new variants, as demonstrated by our cross-neutralization experiments. This characteristic is key for rapidly determining immunity against newly emerging variants. Taken together, the novel assay presented here reduces turnaround time significantly while making use of a highly standardized and sensitive SARS-CoV-2 qRT-PCR method as a readout.

Find related publications in this database (Keywords)
SARS-CoV-2
COVID-19
neutralization assay
neutralizing antibodies
surrogate neutralization assay
cross-neutralization
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