tetano
Editor, Senior Moderator
Biol Pharm Bull
. 2021 Jun 19.
doi: 10.1248/bpb.b21-00387. Online ahead of print.
Improved detection sensitivity of an antigen test for SARS-CoV-2 nucleocapsid proteins with thio-NAD cycling
Yuta Kyosei[SUP] 1 [/SUP], Mayuri Namba[SUP] 1 [/SUP], Sou Yamura[SUP] 1 [/SUP], Satoshi Watabe[SUP] 2 [/SUP], Teruki Yoshimura[SUP] 3 [/SUP], Tadahiro Sasaki[SUP] 4 [/SUP], Tatsuo Shioda[SUP] 4 [/SUP], Etsuro Ito[SUP] 1 2 5 [/SUP]
Affiliations
Abstract
Antigen tests for infectious diseases are inexpensive and easy-to-use, but the limit of detection (LOD) is generally higher than that of polymerase chain reaction (PCR) tests, which are considered the gold standard. In the present study, we combined a sandwich enzyme-linked immunosorbent assay (ELISA) with thionicotinamide-adenine dinucleotide (thio-NAD) cycling to improve the LOD of antigen tests for coronavirus disease 2019 (COVID-19). For recombinant nucleocapsid proteins of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the LOD of our ELISA with thio-NAD cycling was 2.95×10[SUP]-17[/SUP] moles/assay. When UV-irradiated inactive SARS-CoV-2 was used, the minimum detectable virions corresponding to 2.6×10[SUP]4[/SUP] RNA copies/assay were obtained using our ELISA with thio-NAD cycling. The assay volume for each test was 100 μL. The minimum detectable value was smaller than that of the latest antigen test using a fluorescent immunoassay for SARS-CoV-2, indicating the validity of our detection system for COVID-19 diagnosis.
Keywords: antigen test; coronavirus disease 2019; nucleocapsid protein; severe acute respiratory syndrome coronavirus 2; thionicotinamide-adenine dinucleotide cycling; ultrasensitivity.
. 2021 Jun 19.
doi: 10.1248/bpb.b21-00387. Online ahead of print.
Improved detection sensitivity of an antigen test for SARS-CoV-2 nucleocapsid proteins with thio-NAD cycling
Yuta Kyosei[SUP] 1 [/SUP], Mayuri Namba[SUP] 1 [/SUP], Sou Yamura[SUP] 1 [/SUP], Satoshi Watabe[SUP] 2 [/SUP], Teruki Yoshimura[SUP] 3 [/SUP], Tadahiro Sasaki[SUP] 4 [/SUP], Tatsuo Shioda[SUP] 4 [/SUP], Etsuro Ito[SUP] 1 2 5 [/SUP]
Affiliations
- PMID: 34148926
- DOI: 10.1248/bpb.b21-00387
Abstract
Antigen tests for infectious diseases are inexpensive and easy-to-use, but the limit of detection (LOD) is generally higher than that of polymerase chain reaction (PCR) tests, which are considered the gold standard. In the present study, we combined a sandwich enzyme-linked immunosorbent assay (ELISA) with thionicotinamide-adenine dinucleotide (thio-NAD) cycling to improve the LOD of antigen tests for coronavirus disease 2019 (COVID-19). For recombinant nucleocapsid proteins of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the LOD of our ELISA with thio-NAD cycling was 2.95×10[SUP]-17[/SUP] moles/assay. When UV-irradiated inactive SARS-CoV-2 was used, the minimum detectable virions corresponding to 2.6×10[SUP]4[/SUP] RNA copies/assay were obtained using our ELISA with thio-NAD cycling. The assay volume for each test was 100 μL. The minimum detectable value was smaller than that of the latest antigen test using a fluorescent immunoassay for SARS-CoV-2, indicating the validity of our detection system for COVID-19 diagnosis.
Keywords: antigen test; coronavirus disease 2019; nucleocapsid protein; severe acute respiratory syndrome coronavirus 2; thionicotinamide-adenine dinucleotide cycling; ultrasensitivity.