tetano
Editor, Senior Moderator
Biosens Bioelectron
. 2020 Oct 15;171:112715.
doi: 10.1016/j.bios.2020.112715. Online ahead of print.
A novel rapid detection for SARS-CoV-2 spike 1 antigens using human angiotensin converting enzyme 2 (ACE2)
Jong-Hwan Lee[SUP] 1 [/SUP], Minsuk Choi[SUP] 1 [/SUP], Yujin Jung[SUP] 1 [/SUP], Sung Kyun Lee[SUP] 1 [/SUP], Chang-Seop Lee[SUP] 2 [/SUP], Jung Kim[SUP] 1 [/SUP], Jongwoo Kim[SUP] 1 [/SUP], Nam Hoon Kim[SUP] 1 [/SUP], Bum-Tae Kim[SUP] 1 [/SUP], Hong Gi Kim[SUP] 3 [/SUP]
Affiliations
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease 2019 (COVID-19), a newly emerging human infectious disease. Because no specific antiviral drugs or vaccines are available to treat COVID-19, early diagnostics, isolation, and prevention are crucial for containing the outbreak. Molecular diagnostics using reverse transcription polymerase chain reaction (RT-PCR) are the current gold standard for detection. However, viral RNAs are much less stable during transport and storage than proteins such as antigens and antibodies. Consequently, false-negative RT-PCR results can occur due to inadequate collection of clinical specimens or poor handling of a specimen during testing. Although antigen immunoassays are stable diagnostics for detection of past infection, infection progress, and transmission dynamics, no matched antibody pair for immunoassay of SARS-CoV-2 antigens has yet been reported. In this study, we designed and developed a novel rapid detection method for SARS-CoV-2 spike 1 (S1) protein using the SARS-CoV-2 receptor ACE2, which can form matched pairs with commercially available antibodies. ACE2 and S1-mAb were paired with each other for capture and detection in a lateral flow immunoassay (LFIA) that did not cross-react with SARS-CoV Spike 1 or MERS-CoV Spike 1 protein. The SARS-CoV-2 S1 (<5 ng of recombinant proteins/reaction) was detected by the ACE2-based LFIA. The limit of detection of our ACE2-LFIA was 1.86 ? 10[SUP]5[/SUP] copies/mL in the clinical specimen of COVID-19 Patients without no cross-reactivity for nasal swabs from healthy subjects. This is the first study to detect SARS-CoV-2 S1 antigen using an LFIA with matched pair consisting of ACE2 and antibody. Our findings will be helpful to detect the S1 antigen of SARS-CoV-2 from COVID-19 patients.
Keywords: COVID-19; Human ACE2; Lateral flow immunoassay; SARS-CoV-2; Spike antigens.
. 2020 Oct 15;171:112715.
doi: 10.1016/j.bios.2020.112715. Online ahead of print.
A novel rapid detection for SARS-CoV-2 spike 1 antigens using human angiotensin converting enzyme 2 (ACE2)
Jong-Hwan Lee[SUP] 1 [/SUP], Minsuk Choi[SUP] 1 [/SUP], Yujin Jung[SUP] 1 [/SUP], Sung Kyun Lee[SUP] 1 [/SUP], Chang-Seop Lee[SUP] 2 [/SUP], Jung Kim[SUP] 1 [/SUP], Jongwoo Kim[SUP] 1 [/SUP], Nam Hoon Kim[SUP] 1 [/SUP], Bum-Tae Kim[SUP] 1 [/SUP], Hong Gi Kim[SUP] 3 [/SUP]
Affiliations
- PMID: 33099241
- DOI: 10.1016/j.bios.2020.112715
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease 2019 (COVID-19), a newly emerging human infectious disease. Because no specific antiviral drugs or vaccines are available to treat COVID-19, early diagnostics, isolation, and prevention are crucial for containing the outbreak. Molecular diagnostics using reverse transcription polymerase chain reaction (RT-PCR) are the current gold standard for detection. However, viral RNAs are much less stable during transport and storage than proteins such as antigens and antibodies. Consequently, false-negative RT-PCR results can occur due to inadequate collection of clinical specimens or poor handling of a specimen during testing. Although antigen immunoassays are stable diagnostics for detection of past infection, infection progress, and transmission dynamics, no matched antibody pair for immunoassay of SARS-CoV-2 antigens has yet been reported. In this study, we designed and developed a novel rapid detection method for SARS-CoV-2 spike 1 (S1) protein using the SARS-CoV-2 receptor ACE2, which can form matched pairs with commercially available antibodies. ACE2 and S1-mAb were paired with each other for capture and detection in a lateral flow immunoassay (LFIA) that did not cross-react with SARS-CoV Spike 1 or MERS-CoV Spike 1 protein. The SARS-CoV-2 S1 (<5 ng of recombinant proteins/reaction) was detected by the ACE2-based LFIA. The limit of detection of our ACE2-LFIA was 1.86 ? 10[SUP]5[/SUP] copies/mL in the clinical specimen of COVID-19 Patients without no cross-reactivity for nasal swabs from healthy subjects. This is the first study to detect SARS-CoV-2 S1 antigen using an LFIA with matched pair consisting of ACE2 and antibody. Our findings will be helpful to detect the S1 antigen of SARS-CoV-2 from COVID-19 patients.
Keywords: COVID-19; Human ACE2; Lateral flow immunoassay; SARS-CoV-2; Spike antigens.