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

Upsala Journal of Medical Sciences

卷:127

Andris Kazaks¹  Renate Ranka¹  Zanna Rudevica¹  Viktorija Igumnova¹  Ainars Leonciks¹  Dace Skrastina¹  Agnija Kivrane¹  Aigars Reinis²  Anna Zilde²  Svjatoslavs Kistkins²  Elza Elizabete Liepina³ 

[1] Latvian Biomedical Research and Study centre, Ratsupites Street 1, k–1, Riga, LV1067, Latvia;

[2] Pauls Stradins Clinical University Hospital, Pilsonu street 13, Riga, LV1002, Latvia;

[3] Riga Stradins University, Dzirciema Street 16, Riga, LV1007, Latvia;

关键词: lateral flow assay;    elisa;    covid-19;    polyclonal antibodies;    antigen test;    point-of-care testing;   

DOI  :  10.48101/ujms.v127.8207

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown