Temperature-Responsive Liposome-Linked Immunosorbent Assay for the Rapid Detection of SARS-CoV-2 Using Immunoliposomes

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which is the etiological agent of coronavirus disease 2019 (COVID-19), has infected more than 340 million people since the outbreak of the pandemic in 2019, resulting in approximately 55 million deaths. The rapid and effective diagnosis of COVID-19 patients is vital to prevent the spread of the disease. In a previous study, we reported a novel temperature-responsive liposome-linked immunosorbent assay (TLip-LISA) using biotinylated-TLip that exhibited high detection sensitivity for the prostate-specific antigen. Herein, we used immunoglobulin-TLip (IgG-TLip), in which the antibodies were directly conjugated to the liposomal surface to simplify pretreatment procedures and reduce the detection time for SARS-CoV-2. The results indicated that TLip-LISA could detect the recombinant nucleocapsid protein and the nucleocapsid protein in inactivated virus with 20 min incubation time in total, and the limit of detection was calculated to be 2.2 and 1.0 pg/mL, respectively. Therefore, TLip-LISA has high potential to be used in clinic for rapid diagnosis and disease control.

[1]  Sameer Phalke,et al.  SARS-CoV-2 Mutations and Their Impact on Diagnostics, Therapeutics and Vaccines , 2022, Frontiers in Medicine.

[2]  J. Tate,et al.  Performance Characteristics of the Abbott BinaxNOW SARS-CoV-2 Antigen Test in Comparison to Real-Time Reverse Transcriptase PCR and Viral Culture in Community Testing Sites during November 2020 , 2021, Journal of clinical microbiology.

[3]  M. Bewley,et al.  Monoclonal Antibodies to S and N SARS-CoV-2 Proteins as Probes to Assess Structural and Antigenic Properties of Coronaviruses , 2021, Viruses.

[4]  F. Centrone,et al.  The Challenge of Using an Antigen Test as a Screening Tool for SARS-CoV-2 Infection in an Emergency Department: Experience of a Tertiary Care Hospital in Southern Italy , 2021, BioMed research international.

[5]  N. Ohmagari,et al.  Time-course evaluation of the quantitative antigen test for severe acute respiratory syndrome coronavirus 2: The potential contribution to alleviating isolation of COVID-19 patients , 2021, Journal of Infection and Chemotherapy.

[6]  M. Killerby,et al.  Comparison of the SARS-CoV-2 spike protein ELISA and the Abbott Architect SARS-CoV-2 IgG nucleocapsid protein assays for detection of antibodies , 2021, PloS one.

[7]  N. Mach,et al.  A Quantitative ELISA Protocol for Detection of Specific Human IgG against the SARS-CoV-2 Spike Protein , 2021, Vaccines.

[8]  A. Ryo,et al.  Highly specific monoclonal antibodies and epitope identification against SARS-CoV-2 nucleocapsid protein for antigen detection tests , 2021, Cell Reports Medicine.

[9]  M. Boccellino,et al.  Detection of SARS-COV-2 Proteins Using an ELISA Test , 2021, Diagnostics.

[10]  A. Mencacci,et al.  Evaluation of Lumipulse® G SARS-CoV-2 antigen assay automated test for detecting SARS-CoV-2 nucleocapsid protein (NP) in nasopharyngeal swabs for community and population screening , 2021, International Journal of Infectious Diseases.

[11]  Yuta Kyosei,et al.  Antigen tests for COVID-19 , 2021, Biophysics and physicobiology.

[12]  G. Gao,et al.  Viral targets for vaccines against COVID-19 , 2020, Nature reviews. Immunology.

[13]  S. Yagi,et al.  Clinical validation of quantitative SARS-CoV-2 antigen assays to estimate SARS-CoV-2 viral loads in nasopharyngeal swabs , 2020, Journal of Infection and Chemotherapy.

[14]  Y. Kawaoka,et al.  Comparison of Rapid Antigen Tests for COVID-19 , 2020, Viruses.

[15]  K. He,et al.  Foundation and Clinical Evaluation of a New Method for Detecting SARS-CoV-2 Antigen by Fluorescent Microsphere Immunochromatography , 2020, Frontiers in Cellular and Infection Microbiology.

[16]  E. Möncke-Buchner,et al.  Comparison of seven commercial SARS-CoV-2 rapid Point-of-Care Antigen tests , 2020, medRxiv.

[17]  S. Takeoka,et al.  A rapid and highly sensitive biomarker detection platform based on a temperature-responsive liposome-linked immunosorbent assay , 2020, Scientific Reports.

[18]  W. Baumeister,et al.  High-resolution structure and biophysical characterization of the nucleocapsid phosphoprotein dimerization domain from the Covid-19 severe acute respiratory syndrome coronavirus 2 , 2020, Biochemical and Biophysical Research Communications.

[19]  P. Cane,et al.  Analysis of Inactivation of SARS-CoV-2 by Specimen Transport Media, Nucleic Acid Extraction Reagents, Detergents, and Fixatives , 2020, Journal of Clinical Microbiology.

[20]  M. Maejima,et al.  Comparison of automated SARS-CoV-2 antigen test for COVID-19 infection with quantitative RT-PCR using 313 nasopharyngeal swabs, including from seven serially followed patients , 2020, International Journal of Infectious Diseases.

[21]  H. Krumholz,et al.  Extrapulmonary manifestations of COVID-19 , 2020, Nature Medicine.

[22]  James T. Gordy,et al.  The Nucleocapsid Protein of SARS–CoV-2: a Target for Vaccine Development , 2020, Journal of Virology.

[23]  Gheyath K Nasrallah,et al.  Challenges in Laboratory Diagnosis of the Novel Coronavirus SARS-CoV-2 , 2020, Viruses.

[24]  Sairaj Satarker,et al.  Structural Proteins in Severe Acute Respiratory Syndrome Coronavirus-2 , 2020, Archives of Medical Research.

[25]  Daniel S. Chertow,et al.  Sensitivity in Detection of Antibodies to Nucleocapsid and Spike Proteins of Severe Acute Respiratory Syndrome Coronavirus 2 in Patients With Coronavirus Disease 2019 , 2020, The Journal of infectious diseases.

[26]  Fumihiro Kato,et al.  Enhanced isolation of SARS-CoV-2 by TMPRSS2-expressing cells , 2020, Proceedings of the National Academy of Sciences.

[27]  A. M. Leontovich,et al.  The species Severe acute respiratory syndrome-related coronavirus: classifying 2019-nCoV and naming it SARS-CoV-2 , 2020, Nature Microbiology.

[28]  G. Gao,et al.  A Novel Coronavirus from Patients with Pneumonia in China, 2019 , 2020, The New England journal of medicine.

[29]  S. Arai,et al.  Highly cooperative fluorescence switching of self-assembled squaraine dye at tunable threshold temperatures using thermosensitive nanovesicles for optical sensing and imaging , 2019, Scientific Reports.

[30]  M. Ulaşlı,et al.  Nucleocapsid Protein Recruitment to Replication-Transcription Complexes Plays a Crucial Role in Coronaviral Life Cycle , 2019, Journal of Virology.

[31]  Yi Guan,et al.  Genomic Analysis of the Emergence, Evolution, and Spread of Human Respiratory RNA Viruses. , 2016, Annual review of genomics and human genetics.

[32]  R. Hodinka Respiratory RNA Viruses , 2016, Microbiology spectrum.

[33]  P. Masters,et al.  The Molecular Biology of Coronaviruses , 2006, Advances in Virus Research.

[34]  Ben Berkhout,et al.  Human coronavirus NL63 employs the severe acute respiratory syndrome coronavirus receptor for cellular entry , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[35]  I. Kurane,et al.  Immunological detection of severe acute respiratory syndrome coronavirus by monoclonal antibodies. , 2005, Japanese journal of infectious diseases.

[36]  Obi L. Griffith,et al.  The Genome Sequence of the SARS-Associated Coronavirus , 2003, Science.