Development of rapid antigen test prototype for detection of SARS-CoV-2 in saliva samples
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) viru...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Upsala Medical Society
2022-02-01
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| Series: | Upsala Journal of Medical Sciences |
| Subjects: | |
| Online Access: | https://ujms.net/index.php/ujms/article/view/8207/14343 |
| _version_ | 1797719625677406208 |
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| author | Agnija Kivrane Viktorija Igumnova Elza Elizabete Liepina Dace Skrastina Ainars Leonciks Zanna Rudevica Svjatoslavs Kistkins Aigars Reinis Anna Zilde Andris Kazaks Renate Ranka |
| author_facet | Agnija Kivrane Viktorija Igumnova Elza Elizabete Liepina Dace Skrastina Ainars Leonciks Zanna Rudevica Svjatoslavs Kistkins Aigars Reinis Anna Zilde Andris Kazaks Renate Ranka |
| author_sort | Agnija Kivrane |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-12T09:07:43Z |
| format | Article |
| id | doaj.art-4115387007de4d0683ceedfcfde7ce46 |
| institution | Directory Open Access Journal |
| issn | 0300-9734 2000-1967 |
| language | English |
| last_indexed | 2024-03-12T09:07:43Z |
| publishDate | 2022-02-01 |
| publisher | Upsala Medical Society |
| record_format | Article |
| series | Upsala Journal of Medical Sciences |
| spelling | doaj.art-4115387007de4d0683ceedfcfde7ce462023-09-02T15:11:31ZengUpsala Medical SocietyUpsala Journal of Medical Sciences0300-97342000-19672022-02-011271910.48101/ujms.v127.82078207Development of rapid antigen test prototype for detection of SARS-CoV-2 in saliva samplesAgnija Kivrane0Viktorija Igumnova1Elza Elizabete Liepina2Dace Skrastina3Ainars Leonciks4Zanna Rudevica5Svjatoslavs Kistkins6Aigars Reinis7Anna Zilde8Andris Kazaks9Renate Ranka10Latvian Biomedical Research and Study centre, Ratsupites Street 1, k–1, Riga, LV1067, LatviaLatvian Biomedical Research and Study centre, Ratsupites Street 1, k–1, Riga, LV1067, LatviaRiga Stradins University, Dzirciema Street 16, Riga, LV1007, LatviaLatvian Biomedical Research and Study centre, Ratsupites Street 1, k–1, Riga, LV1067, LatviaLatvian Biomedical Research and Study centre, Ratsupites Street 1, k–1, Riga, LV1067, LatviaLatvian Biomedical Research and Study centre, Ratsupites Street 1, k–1, Riga, LV1067, LatviaPauls Stradins Clinical University Hospital, Pilsonu street 13, Riga, LV1002, LatviaPauls Stradins Clinical University Hospital, Pilsonu street 13, Riga, LV1002, LatviaPauls Stradins Clinical University Hospital, Pilsonu street 13, Riga, LV1002, LatviaLatvian Biomedical Research and Study centre, Ratsupites Street 1, k–1, Riga, LV1067, LatviaLatvian Biomedical Research and Study centre, Ratsupites Street 1, k–1, Riga, LV1067, LatviaBackground: 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.https://ujms.net/index.php/ujms/article/view/8207/14343lateral flow assayelisacovid-19polyclonal antibodiesantigen testpoint-of-care testing |
| spellingShingle | Agnija Kivrane Viktorija Igumnova Elza Elizabete Liepina Dace Skrastina Ainars Leonciks Zanna Rudevica Svjatoslavs Kistkins Aigars Reinis Anna Zilde Andris Kazaks Renate Ranka Development of rapid antigen test prototype for detection of SARS-CoV-2 in saliva samples Upsala Journal of Medical Sciences lateral flow assay elisa covid-19 polyclonal antibodies antigen test point-of-care testing |
| title | Development of rapid antigen test prototype for detection of SARS-CoV-2 in saliva samples |
| title_full | Development of rapid antigen test prototype for detection of SARS-CoV-2 in saliva samples |
| title_fullStr | Development of rapid antigen test prototype for detection of SARS-CoV-2 in saliva samples |
| title_full_unstemmed | Development of rapid antigen test prototype for detection of SARS-CoV-2 in saliva samples |
| title_short | Development of rapid antigen test prototype for detection of SARS-CoV-2 in saliva samples |
| title_sort | development of rapid antigen test prototype for detection of sars cov 2 in saliva samples |
| topic | lateral flow assay elisa covid-19 polyclonal antibodies antigen test point-of-care testing |
| url | https://ujms.net/index.php/ujms/article/view/8207/14343 |
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