SalivaSTAT: Direct-PCR and Pooling of Saliva Samples Collected in Healthcare and Community Setting for SARS-CoV-2 Mass Surveillance
Authors
Sahajpal, Nikhil S.Mondal, Ashis K.
Ananth, Sudha
Njau, Allan
Ahluwalia, Pankaj
Newnam, Gary
Lozoya-Colinas, Adriana
Hud, Nicholas V.
Kota, Vamsi
Ross, Ted M.
Reid, Michelle D.
Fulzele, Sadanand
Chaubey, Alka
Hegde, Madhuri
Rojiani, Amyn M.
Kolhe, Ravindra
Issue Date
2021-05-19
Metadata
Show full item recordAbstract
Objectives: Limitations of widespread current COVID-19 diagnostic testing exist in both the pre-analytical and analytical stages. To alleviate these limitations, we developed a universal saliva processing protocol (SalivaSTAT) that would enable an extraction-free RT-PCR test using commercially available RT-PCR kits. Methods: We optimized saliva collection devices, heat-shock treatment, and homogenization. Saliva samples (879) previously tested using the FDA-EUA method were reevaluated with the optimized SalivaSTAT protocol using two widely available commercial RT-PCR kits. A five-sample pooling strategy was evaluated as per FDA guidelines. Results: Saliva collection (done without any media) showed performance comparable to that of the FDA-EUA method. The SalivaSTAT protocol was optimized by incubating saliva samples at 95 ◦C for 30-min and homogenization, followed by RT-PCR assay. The clinical sample evaluation of 630 saliva samples using the SalivaSTAT protocol with PerkinElmer (600-samples) and CDC (30-samples) RT-PCR assay achieved positive (PPA) and negative percent agreements (NPAs) of 95.0% and 100%, respectively. The LoD was established as ~60–180 copies/mL by absolute quantification. Furthermore, a five-samplepooling evaluation using 250 saliva samples achieved a PPA and NPA of 92% and 100%, respectively. Conclusion: We have optimized an extraction-free RT-PCR assay for saliva samples that demonstrates comparable performance to FDA-EUA assay (Extraction and RT-PCR).Affiliation
Department of Pathology; Department of Medicine; Center for Healthy Agingae974a485f413a2113503eed53cd6c53
10.3390/diagnostics11050904
Scopus Count
Collections
Except where otherwise noted, this item's license is described as https://creativecommons.org/licenses/by/4.0/