{"doi":"10.3390/diagnostics11050904","title":"SalivaSTAT: Direct-PCR and Pooling of Saliva Samples Collected in Healthcare and Community Setting for SARS-CoV-2 Mass Surveillance","abstract":"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-sample-pooling 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).","journal":"Diagnostics","year":2021,"id":178057,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":19,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9521,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2021-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":703901,"name":"Ashis K. Mondal","orcid":"0000-0003-3826-9489","position":1,"is_corresponding":false},{"id":703902,"name":"Sudha Ananth","orcid":"0000-0001-9951-5474","position":2,"is_corresponding":false},{"id":703903,"name":"Allan Njau","orcid":"0000-0002-3566-0629","position":3,"is_corresponding":false},{"id":703904,"name":"Pankaj Ahluwalia","orcid":"0000-0003-0060-6152","position":4,"is_corresponding":false},{"id":723797,"name":"Gary Newnam","orcid":null,"position":5,"is_corresponding":false},{"id":723798,"name":"Adriana Lozoya-Colinas","orcid":null,"position":6,"is_corresponding":false},{"id":337557,"name":"Nicholas V. Hud","orcid":"0000-0001-7711-6472","position":7,"is_corresponding":false},{"id":703905,"name":"Vamsi Kota","orcid":"0000-0002-5290-9289","position":8,"is_corresponding":false},{"id":245069,"name":"Ted M. Ross","orcid":"0000-0003-1947-7469","position":9,"is_corresponding":false},{"id":416161,"name":"Michelle D. Reid","orcid":"0000-0002-3812-4098","position":10,"is_corresponding":false},{"id":264778,"name":"Sadanand Fulzele","orcid":"0000-0002-3510-6759","position":11,"is_corresponding":false},{"id":703909,"name":"Alka Chaubey","orcid":"0000-0002-0914-9237","position":12,"is_corresponding":false},{"id":55849,"name":"Madhuri Hegde","orcid":"0000-0002-4500-0368","position":13,"is_corresponding":false},{"id":703910,"name":"Amyn M. Rojiani","orcid":"0000-0001-5621-3724","position":14,"is_corresponding":false},{"id":264782,"name":"Ravindra Kolhe","orcid":"0000-0002-8283-2403","position":15,"is_corresponding":false},{"id":723796,"name":"Nikhil S. Sahajpal","orcid":null,"position":0,"is_corresponding":true}],"reference_count":28,"raw_metadata":null,"created_at":"2026-07-18T23:47:36.112170Z","pmid":"34069462","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}