Development of rapid antigen test prototype for detection of SARS-CoV-2 in saliva samples

  • Agnija Kivrane Latvian Biomedical Research and Study centre, Ratsupites Street 1, k–1, Riga, LV1067, Latvia; and Riga Stradins University, Dzirciema Street 16, Riga, LV1007, Latvia https://orcid.org/0000-0002-8284-2011
  • Viktorija Igumnova Latvian Biomedical Research and Study centre, Ratsupites Street 1, k–1, Riga, LV1067, Latvia; and Riga Stradins University, Dzirciema Street 16, Riga, LV1007, Latvia https://orcid.org/0000-0002-5028-2718
  • Elza Elizabete Liepina Riga Stradins University, Dzirciema Street 16, Riga, LV1007, Latvia
  • Dace Skrastina Latvian Biomedical Research and Study centre, Ratsupites Street 1, k–1, Riga, LV1067, Latvia https://orcid.org/0000-0001-5836-6441
  • Ainars Leonciks Latvian Biomedical Research and Study centre, Ratsupites Street 1, k–1, Riga, LV1067, Latvia
  • Zanna Rudevica Latvian Biomedical Research and Study centre, Ratsupites Street 1, k–1, Riga, LV1067, Latvia
  • Svjatoslavs Kistkins Pauls Stradins Clinical University Hospital, Pilsonu street 13, Riga, LV1002, Latvia
  • Aigars Reinis Pauls Stradins Clinical University Hospital, Pilsonu street 13, Riga, LV1002, Latvia https://orcid.org/0000-0002-3671-1213
  • Anna Zilde Pauls Stradins Clinical University Hospital, Pilsonu street 13, Riga, LV1002, Latvia
  • Andris Kazaks Latvian Biomedical Research and Study centre, Ratsupites Street 1, k–1, Riga, LV1067, Latvia https://orcid.org/0000-0003-4964-0984
  • Renate Ranka Latvian Biomedical Research and Study centre, Ratsupites Street 1, k–1, Riga, LV1067, Latvia; and Riga Stradins University, Dzirciema Street 16, Riga, LV1007, Latvia https://orcid.org/0000-0002-3716-7950
Keywords: Lateral flow assay, ELISA, COVID-19, polyclonal antibodies, antigen test, point-of-care testing

Abstract

Background: The development of easy-to-perform diagnostic methods is highly important for detecting current coronavirus disease (COVID-19). This pilot study aimed at developing a lateral flow assay (LFA)-based test prototype to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus in saliva samples.

Methods: Mice were immunized using the recombinant receptor-binding domain (rRBD) of SARS-CoV-2 virus spike protein. The combinations of the obtained mouse anti-receptor-binding domain (RBD) polyclonal antibodies (PAbs) and several commercial antibodies directed against the SARS-CoV-2 spike protein were used for enzyme-linked immunosorbent assay (ELISA) to select antibody pairs for LFA. The antibody pairs were tested in a LFA format using saliva samples from individuals with early SARS-CoV-2 infection (n = 9). The diagnostic performance of the developed LFA was evaluated using saliva samples from hospitalized COVID-19 patients (n = 111); the median time from the onset of symptoms to sample collection was 10 days (0–24 days, interquartile range (IQR): 7–13). The reverse transcription-polymerase chain reaction (rRT-PCR) was used as a reference method.

Results: Based on ELISA and preliminary LFA results, a combination of mouse anti-RBD PAbs (capture antibody) and rabbit anti-spike PAbs (detection antibody) was chosen for clinical analysis of sample. When compared with rRT-PCR results, LFA exhibited 26.5% sensitivity, 58.1% specificity, 50.0% positive prediction value (PPV), 33.3% negative prediction value (NPV), and 38.7% diagnostic accuracy. However, there was a reasonable improvement in assay specificity (85.7%) and PPV (91.7%) when samples were stratified based on the sampling time.

Conclusion: The developed LFA assay demonstrated a potential of SARS-CoV-2 detection in saliva samples. Further technical assay improvements should be made to enhance diagnostic performance followed by a validation study in a larger cohort of both asymptomatic and symptomatic patients in the early stage of infection.

Downloads

Download data is not yet available.

References


  1. Mach F, Baigent C, Catapano AL, Koskinas KC, Casula M, Badimon L, et al. 2019 ESC/EAS guidelines for the management of dyslipidaemias: lipid modification to reduce cardiovascular risk. Eur Heart J. 2020;14:111–88. doi: 10.1093/eurheartj/ehz455

  2. American Heart Association. Cardiovascular disease: a costly burden for America. Projections through 2035. Available from: http://www.heart.org/idc/groups/heart-public/@wcm/@adv/documents/downloadable/ucm_491543.pdf (accessed January 2022).

  3. Mendis S. Global progress in prevention of cardiovascular disease. Cardiovasc Diagn Ther. 2017;7:S32–8. doi: 10.21037/cdt.2017.03.06

  4. Townsend N, Wilson L, Bhatnagar P, Wickramasinghe K, Rayner M, Nichols M. Cardiovascular disease in Europe: epidemiological update 2016. Eur Heart J. 2016;37:3232–45. doi: 10.1093/eurheartj/ehw334

  5. Bhatnagar D, Soran H, Durrington PN. Hypercholesterolaemia and its management. BMJ. 2008;337:a993. doi: 10.1136/bmj.a993

  6. Roth GA, Fihn SD, Mokdad AH, Aekplakorn W, Hasegawa T, Lim SS. High total serum cholesterol, medication coverage and therapeutic control: an analysis of national health examination survey data from eight countries. Bull World Health Organ. 2011;89:92–101. doi: 10.2471/BLT.10.079947

  7. Journath G, Hambraeus K, Hagstrom E, Pettersson B, Lothgren M. Predicted impact of lipid lowering therapy on cardiovascular and economic outcomes of Swedish atherosclerotic cardiovascular disease guideline. BMC Cardiovasc Disord. 2017;17:224. doi: 10.1186/s12872-017-0659-2

  8. Piepoli MF, Hoes AW, Agewall S, Albus C, Brotons C, Catapano AL, et al. [2016 European guidelines on cardiovascular disease prevention in clinical practice. The Sixth Joint Task Force of the European Society of Cardiology and Other Societies on Cardiovascular Disease Prevention in Clinical Practice (constituted by representatives of 10 societies and by invited experts. Developed with the special contribution of the European Association for Cardiovascular Prevention & Rehabilitation]. G Ital Cardiol (Rome). 2017;18:547–612.

  9. Kotseva K, De Backer G, De Bacquer D, Ryden L, Hoes A, Grobbee D, et al. Lifestyle and impact on cardiovascular risk factor control in coronary patients across 27 countries: results from the European Society of Cardiology ESC-EORP EUROASPIRE V registry. Eur J Prev Cardiol. 2019;26:824–35. doi: 10.1177/2047487318825350

  10. Schubert J, Lindahl B, Melhus H, Renlund H, Leosdottir M, Yari A, et al. Low-density lipoprotein cholesterol reduction and statin intensity in myocardial infarction patients and major adverse outcomes: a Swedish nationwide cohort study. Eur Heart J. 2021;42:243–52. doi: 10.1093/eurheartj/ehaa1011

  11. Khalaf K, Johnell K, Austin PC, Tyden P, Midlov P, Perez-Vicente R, et al. Low adherence to statin treatment during the first year after an acute myocardial infarction is associated with increased second year mortality risk – an inverse probability of treatment weighted study on 54,872 patients. Eur Heart J Cardiovasc Pharmacother. 2021;7:141–7. doi: 10.1093/ehjcvp/pvaa010

  12. Karlsson SA, Eliasson B, Franzen S, Miftaraj M, Svensson AM, Andersson Sundell K. Risk of cardiovascular event and mortality in relation to refill and guideline adherence to lipid-lowering medications among patients with type 2 diabetes mellitus in Sweden. BMJ Open Diabetes Res Care. 2019;7:e000639. doi: 10.1136/bmjdrc-2018-000639

  13. Nilsson G, Samuelsson E, Soderstrom L, Mooe T. Low use of statins for secondary prevention in primary care: a survey in a northern Swedish population. BMC Fam Pract. 2016;17:110. doi: 10.1186/s12875-016-0505-0

  14. Kotseva K, Wood D, De Bacquer D, De Backer G, Ryden L, Jennings C, et al. EUROASPIRE IV: a European Society of Cardiology survey on the lifestyle, risk factor and therapeutic management of coronary patients from 24 European countries. Eur J Prev Cardiol. 2016;23:636–48. doi: 10.1177/2047487315569401

  15. Modig S, Hoglund P, Troein M, Midlov P. GP’s adherence to guidelines for cardiovascular disease among elderly: a quality development study. ScientificWorldJournal. 2012;2012:767892. doi: 10.1100/2012/767892

  16. Hallberg S, Banefelt J, Fox KM, Mesterton J, Johansson G, Levin LA, et al. Lipid-lowering treatment patterns in patients with new cardiovascular events – estimates from population-based register data in Sweden. Int J Clin Pract. 2016;70:222–8. doi: 10.1111/ijcp.12769

  17. Ludvigsson JF, Andersson E, Ekbom A, Feychting M, Kim JL, Reuterwall C, et al. External review and validation of the Swedish national inpatient register. BMC Public Health. 2011;11:450. doi: 10.1186/1471-2458-11-450

  18. Ludvigsson JF, Otterblad-Olausson P, Pettersson BU, Ekbom A. The Swedish personal identity number: possibilities and pitfalls in healthcare and medical research. Eur J Epidemiol. 2009;24:659–67. doi: 10.1007/s10654-009-9350-y

  19. Wettermark B, Hammar N, Fored CM, Leimanis A, Olausson PO, Bergman U, et al. The new Swedish Prescribed Drug Register – opportunities for pharmacoepidemiological research and experience from the first six months. Pharmacoepidemiol Drug Saf. 2007;16:726–35. doi: 10.1002/pds.1294

  20. Perk J, De Backer G, Gohlke H, Graham I, Reiner Z, Verschuren M, et al. European guidelines on cardiovascular disease prevention in clinical practice (version 2012). The fifth Joint Task Force of the European Society of Cardiology and other societies on cardiovascular disease prevention in clinical practice (constituted by representatives of nine societies and by invited experts). Eur Heart J. 2012;33:1635–701. doi: 10.1093/eurheartj/ehs092

  21. Läkemedelsverket. Att förebygga aterosklerotisk hjärt-kärlsjukdom med läkemedel – behandlingsrekommendation. 2014;25: 20–33. Available from: https://www.lakemedelsverket.se/sv/behandling-och-forskrivning/behandlingsrekommendationer/sok-behandlingsrekommendationer/lakemedel-for-att-forebygga-aterosklerotisk-hjart-karlsjukdom--behandlingsrekommendation (accessed January 2022).

  22. Stone NJ, Robinson JG, Lichtenstein AH, Bairey Merz CN, Blum CB, Eckel RH, et al. 2013 ACC/AHA guideline on the treatment of blood cholesterol to reduce atherosclerotic cardiovascular risk in adults: a report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines. Circulation. 2014;129:S1–45. doi: 10.1161/01.cir.0000437738.63853.7a

  23. Reiner Z, De Backer G, Fras Z, Kotseva K, Tokgozoglu L, Wood D, et al. Lipid lowering drug therapy in patients with coronary heart disease from 24 European countries – findings from the EUROASPIRE IV survey. Atherosclerosis. 2016;246:243–50. doi: 10.1016/j.atherosclerosis.2016.01.018

  24. Steg PG, James SK, Atar D, Badano LP, Blomstrom-Lundqvist C, Borger MA, et al. ESC Guidelines for the management of acute myocardial infarction in patients presenting with ST-segment elevation. Eur Heart J. 2012;33:2569–619. doi: 10.1093/eurheartj/ehs215

  25. Montalescot G, Sechtem U, Achenbach S, Andreotti F, Arden C, Budaj A, et al. 2013 ESC guidelines on the management of stable coronary artery disease: the Task Force on the management of stable coronary artery disease of the European Society of Cardiology. Eur Heart J. 2013;34:2949–3003. doi: 10.1093/eurheartj/eht296

  26. Khunti K, Danese MD, Kutikova L, Catterick D, Sorio-Vilela F, Gleeson M, et al. Association of a combined measure of adherence and treatment intensity with cardiovascular outcomes in patients with atherosclerosis or other cardiovascular risk factors treated with statins and/or ezetimibe. JAMA Netw Open. 2018;1:e185554. doi: 10.1001/jamanetworkopen.2018.5554

  27. Ahrens I, Khachatryan A, Monga B, Dornstauder E, Sidelnikov E. Association of treatment intensity and adherence to lipid-lowering therapy with major adverse cardiovascular events among post-MI patients in Germany. Adv Ther. 2021;38:2532–41. doi: 10.1007/s12325-021-01697-8

  28. Schiele F, Quignot N, Khachatryan A, Gusto G, Villa G, Kahangire D, et al. Clinical impact and room for improvement of intensity and adherence to lipid lowering therapy: five years of clinical follow-up from 164,565 post-myocardial infarction patients. Int J Cardiol. 2021;332:22–8. doi: 10.1016/j.ijcard.2021.03.007

  29. Lindh M, Banefelt J, Fox KM, Hallberg S, Tai MH, Eriksson M, et al. Cardiovascular event rates in a high atherosclerotic cardiovascular disease risk population: estimates from Swedish population-based register data. Eur Heart J Qual Care Clin Outcomes. 2019;5:225–32. doi: 10.1093/ehjqcco/qcy058

  30. Banefelt J, Lindh M, Svensson MK, Eliasson B, Tai M. Statin dose titration patterns and subsequent major cardiovascular events in very high-risk patients – estimates from Swedish population-based registry data. Eur Heart J Qual Care Clin Outcomes. 2020;6:323–31. doi: 10.1093/ehjqcco/qcaa023

Published
2022-02-25
How to Cite
KivraneA., IgumnovaV., LiepinaE. E., SkrastinaD., LeonciksA., RudevicaZ., KistkinsS., ReinisA., ZildeA., KazaksA., & RankaR. (2022). Development of rapid antigen test prototype for detection of SARS-CoV-2 in saliva samples. Upsala Journal of Medical Sciences, 127(1). https://doi.org/10.48101/ujms.v127.8207