tetano
Editor, Senior Moderator
BMC Immunol
. 2023 Sep 26;24(1):34.
doi: 10.1186/s12865-023-00567-y. The development of a highly sensitive and quantitative SARS-CoV-2 rapid antigen test applying newly developed monoclonal antibodies to an automated chemiluminescent flow-through membrane immunoassay device
Kengo Nishimura[SUP] 1 [/SUP], Hiroaki Kitazawa[SUP] 2 [/SUP], Takashi Kawahata[SUP] 2 [/SUP], Kosuke Yuhara[SUP] 2 [/SUP], Takahiro Masuya[SUP] 2 [/SUP], Toshihiro Kuroita[SUP] 3 [/SUP], Kentarou Waki[SUP] 4 [/SUP], Seiichi Koike[SUP] 5 [/SUP], Masaharu Isobe[SUP] 5 [/SUP], Nobuyuki Kurosawa[SUP] 6 [/SUP]
Affiliations
Background: Rapid and accurate diagnosis of individuals with SARS-CoV-2 infection is an effective way to prevent and control the spread of COVID-19. Although the detection of SARS-CoV-2 viral RNA by RT-qPCR is the gold standard for COVID-19 testing, the use of antigen-detecting rapid diagnostic tests (Ag-RDTs) is emerging as a complementary surveillance tool as Omicron case numbers skyrocket worldwide. However, the results from Ag-RDTs are less accurate in individuals with low viral loads.
Results: To develop a highly sensitive and accurate Ag-RDT, 90 monoclonal antibodies were raised from guinea pigs immunized with SARS CoV-2 nucleocapsid protein (CoV-2-NP). By applying a capture antibody recognizing the structural epitope of the N-terminal domain of CoV-2-NP and a detection antibody recognizing the C-terminal tail of CoV-2-NP to an automated chemiluminescence flow-through membrane immunoassay device, we developed a novel Ag-RDT, CoV-2-POCube. The CoV-2-POCube exclusively recognizes CoV-2-NP variants but not the nucleocapsid proteins of other human coronaviruses. The CoV-2-POCube achieved a limit of detection sensitivity of 0.20 ~ 0.66 pg/mL of CoV-2-NPs, demonstrating more than 100 times greater sensitivity than commercially available SARS-CoV-2 Ag-RDTs.
Conclusions: CoV-2-POCube has high analytical sensitivity and can detect SARS-CoV-2 variants in 15 min without observing the high-dose hook effect, thus meeting the need for early SARS-CoV-2 diagnosis with lower viral load. CoV-2-POCube is a promising alternative to currently available diagnostic devices for faster clinical decision making in individuals with suspected COVID-19 in resource-limited settings.
Keywords: COVID-19; Lateral flow immunochromatography; Monoclonal antibody; Omicron; Point-of-care test; Rapid antigen test; SARS-CoV-2.
. 2023 Sep 26;24(1):34.
doi: 10.1186/s12865-023-00567-y. The development of a highly sensitive and quantitative SARS-CoV-2 rapid antigen test applying newly developed monoclonal antibodies to an automated chemiluminescent flow-through membrane immunoassay device
Kengo Nishimura[SUP] 1 [/SUP], Hiroaki Kitazawa[SUP] 2 [/SUP], Takashi Kawahata[SUP] 2 [/SUP], Kosuke Yuhara[SUP] 2 [/SUP], Takahiro Masuya[SUP] 2 [/SUP], Toshihiro Kuroita[SUP] 3 [/SUP], Kentarou Waki[SUP] 4 [/SUP], Seiichi Koike[SUP] 5 [/SUP], Masaharu Isobe[SUP] 5 [/SUP], Nobuyuki Kurosawa[SUP] 6 [/SUP]
Affiliations
- PMID: 37752417
- DOI: 10.1186/s12865-023-00567-y
Background: Rapid and accurate diagnosis of individuals with SARS-CoV-2 infection is an effective way to prevent and control the spread of COVID-19. Although the detection of SARS-CoV-2 viral RNA by RT-qPCR is the gold standard for COVID-19 testing, the use of antigen-detecting rapid diagnostic tests (Ag-RDTs) is emerging as a complementary surveillance tool as Omicron case numbers skyrocket worldwide. However, the results from Ag-RDTs are less accurate in individuals with low viral loads.
Results: To develop a highly sensitive and accurate Ag-RDT, 90 monoclonal antibodies were raised from guinea pigs immunized with SARS CoV-2 nucleocapsid protein (CoV-2-NP). By applying a capture antibody recognizing the structural epitope of the N-terminal domain of CoV-2-NP and a detection antibody recognizing the C-terminal tail of CoV-2-NP to an automated chemiluminescence flow-through membrane immunoassay device, we developed a novel Ag-RDT, CoV-2-POCube. The CoV-2-POCube exclusively recognizes CoV-2-NP variants but not the nucleocapsid proteins of other human coronaviruses. The CoV-2-POCube achieved a limit of detection sensitivity of 0.20 ~ 0.66 pg/mL of CoV-2-NPs, demonstrating more than 100 times greater sensitivity than commercially available SARS-CoV-2 Ag-RDTs.
Conclusions: CoV-2-POCube has high analytical sensitivity and can detect SARS-CoV-2 variants in 15 min without observing the high-dose hook effect, thus meeting the need for early SARS-CoV-2 diagnosis with lower viral load. CoV-2-POCube is a promising alternative to currently available diagnostic devices for faster clinical decision making in individuals with suspected COVID-19 in resource-limited settings.
Keywords: COVID-19; Lateral flow immunochromatography; Monoclonal antibody; Omicron; Point-of-care test; Rapid antigen test; SARS-CoV-2.