A novel rapid detection for SARS-CoV-2 spike 1 antigens using human angiotensin converting enzyme 2 (ACE2)

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 5 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.

Internet of medical things (IoMT)-integrated biosensors for point-of-care testing of infectious diseases

“…The proposed sensor was used as multiplex POCT for simultaneous detection of MERS-CoV, HPV, and Mycobacterium. Recently, developed LFA against SARS-CoV-2 exploited the binding affinity of COVID-19 structural protein S1 to angiotensin-converting enzyme 2 (ACE2) domain ( Lee et al, 2020 ). LFA identify matched pair at 1.86 × 105 copies/mL LOD and specifically without any cross reactivity to other corona viruses like SARS and MERS.…”

Trends in biosensing platforms for SARS-CoV-2 detection: A critical appraisal against standard detection tools

“…Both the lateral flow assays and ELISA methods can be used for the detection of antigens. Kim’s research group developed a novel detection method where they used ACE2 receptor to detect SARS-CoV-2 spike 1 (S1) protein [ 46 ]. The receptor is further paired with commercially available antibodies and this pair was captured and detected using lateral flow immunoassay.…”

New insights into application of nanoparticles in the diagnosis and screening of novel coronavirus (SARS-CoV-2)

“…An innovative idea for rapid and sensitive detection of the SARS-CoV-2 spike-1 (S1) protein has been introduced that is the use of the SARS-CoV-2 receptor ACE-2 protein matched pairs with commercially available antibodies [ 54 ]. ACE-2 and monoclonal antibody against S1 protein (S1-mAb) were paired with each other for detection in a lateral flow immunoassay (LFIA).…”

“…Subsequently, the SARS-CoV-2 S1 protein was recognized in the ACE2-based LFIA system. The limit of detection (LOD) was found to be about 1.86 × 10 5 copies/mL in clinical specimens from COVID-19 patients without any cross-reactivity for nasal swabs [ 54 ]. Owing to all these, it can be said that the biosensors designed based on the nanotechnology are rapid/easy to use, specific in nature without cross reaction.…”