Methods to evaluate the impact of SARS-CoV-2 nucleocapsid mutations on antigen detection by rapid diagnostic tests
BIOTECHNIQUES(2022)
Abstract
Mutations in the nucleocapsid of SARS-CoV-2may interfere with antigen detection by diagnostic tests. We used several methods to evaluate the effect of various SARS-CoV-2 nucleocapsid mutations on the performance of the Panbio (TM) and BinaxNOW (TM) lateral flow rapid antigen tests and a prototype high-throughput immunoassay that utilizes Panbio antibodies. Variant detection was also evaluated by immunoblot and BIAcore (TM) assay. A panel of 23 recombinant nucleocapsid antigens (rAgs) were produced that included mutations found in circulating SARS-CoV-2 variants, including variants of concern. All mutant rAgs were detected by all assays, at a sensitivity equivalent to wild-type control (Wuhan strain). Thus, using a rAg approach, we found that the SARS-CoV-2 nucleocapsid mutations examined do not directly impact antigen detection or antigen assay performance. METHOD SUMMARY Expression, purification and testing of wild-type and mutant recombinant SARS-CoV-2 nucleocapsid antigens by the lateral flow SARS-CoV-2 antigen tests BinaxNOWT and Panbio (TM), an ARCHITECTR (R) SARS-CoV-2 antigen prototype automated immunoassay, immunoblot and BIAcore (TM) (surface plasmon resonance) activity assays.
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Key words
antigen,BinaxNOW,immunoassay,lateral flow assay,nucleocapsid,Panbio,recombinant protein,SARS-CoV-2,variant
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