tetano
Editor, Senior Moderator
Ups J Med Sci
. 2022 Feb 25;127.
doi: 10.48101/ujms.v127.8207. eCollection 2022.
Development of rapid antigen test prototype for detection of SARS-CoV-2 in saliva samples
Agnija Kivrane[SUP] 1 2 [/SUP], Viktorija Igumnova[SUP] 1 2 [/SUP], Elza Elizabete Liepina[SUP] 2 [/SUP], Dace Skrastina[SUP] 1 [/SUP], Ainars Leonciks[SUP] 1 [/SUP], Zanna Rudevica[SUP] 1 [/SUP], Svjatoslavs Kistkins[SUP] 3 [/SUP], Aigars Reinis[SUP] 3 [/SUP], Anna Zilde[SUP] 3 [/SUP], Andris Kazaks[SUP] 1 [/SUP], Renate Ranka[SUP] 1 2 [/SUP]
Affiliations
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.
Keywords: COVID-19; ELISA; Lateral flow assay; antigen test; point-of-care testing; polyclonal antibodies.
. 2022 Feb 25;127.
doi: 10.48101/ujms.v127.8207. eCollection 2022.
Development of rapid antigen test prototype for detection of SARS-CoV-2 in saliva samples
Agnija Kivrane[SUP] 1 2 [/SUP], Viktorija Igumnova[SUP] 1 2 [/SUP], Elza Elizabete Liepina[SUP] 2 [/SUP], Dace Skrastina[SUP] 1 [/SUP], Ainars Leonciks[SUP] 1 [/SUP], Zanna Rudevica[SUP] 1 [/SUP], Svjatoslavs Kistkins[SUP] 3 [/SUP], Aigars Reinis[SUP] 3 [/SUP], Anna Zilde[SUP] 3 [/SUP], Andris Kazaks[SUP] 1 [/SUP], Renate Ranka[SUP] 1 2 [/SUP]
Affiliations
- PMID: 35284045
- PMCID: PMC8886438
- DOI: 10.48101/ujms.v127.8207
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.
Keywords: COVID-19; ELISA; Lateral flow assay; antigen test; point-of-care testing; polyclonal antibodies.