{"doi":"10.1101/2021.05.06.21256733","title":"RT-qPCR-based tests for SARS-CoV-2 detection in pooled saliva samples for massive population screening to monitor epidemics","abstract":"Abstract Swab, quantitative, reverse transcription polymerase chain reaction (RT-qPCR) tests remain the gold standard of diagnostics of SARS-CoV-2 infections. However, these tests are costly and time-consuming, and swabbing limits their throughput. We developed a 3-gene, seminested RT-qPCR test with SYBR green-based detection, optimized for testing pooled saliva samples for high-throughput diagnostics of epidemic-affected populations. The proposed two-tier approach depends on decentralized self-collection of saliva samples, pooling, 1 st -tier testing with the mentioned highly sensitive screening test and subsequent 2 nd -tier testing of individual samples from positive pools with the in vitro diagnostic (IVD) test. The screening test was able to detect 5 copies of the viral genome in 10 µl of isolated RNA with 50% probability and 18.8 copies with 95% probability and reached Ct values that were highly linearly RNA concentration-dependent. In the side-by-side comparison (testing artificial pooled samples), the screening test attained slightly better results than the commercially available IVD-certified RT-qPCR diagnostic test (100% specificity and 89.8% sensitivity vs. 100% and 73.5%, respectively). Testing of 1475 individual clinical samples pooled in 374 pools of 4 revealed 0.8% false positive pools and no false negative pools. In weekly prophylactic testing of 113 people within 6 months, a two-tier testing approach enabled the detection of 18 infected individuals, including several asymptomatic individuals, with a fraction of the costs of individual RT-PCR testing.","journal":"medRxiv","year":2021,"id":224293,"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":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.959,"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":826300,"name":"Aurelia Walczak‐Drzewiecka","orcid":"0000-0001-9519-8774","position":1,"is_corresponding":false},{"id":826735,"name":"Jolanta Witaszewska","orcid":null,"position":2,"is_corresponding":false},{"id":826301,"name":"Ewelina Wójcik","orcid":"0000-0002-9743-9268","position":3,"is_corresponding":false},{"id":826302,"name":"Arkadiusz Guziński","orcid":"0000-0001-5155-9806","position":4,"is_corresponding":false},{"id":826736,"name":"Bogumił Zimoń","orcid":null,"position":5,"is_corresponding":false},{"id":826303,"name":"Rafał Matusiak","orcid":"0000-0002-2796-610X","position":6,"is_corresponding":false},{"id":826304,"name":"Joanna Kazimierczak","orcid":"0000-0002-8667-7949","position":7,"is_corresponding":false},{"id":826305,"name":"Maciej Borowiec","orcid":"0000-0003-2812-5676","position":8,"is_corresponding":false},{"id":826306,"name":"Katarzyna Kania","orcid":"0000-0002-0673-596X","position":9,"is_corresponding":false},{"id":826307,"name":"Edyta Paradowska","orcid":"0000-0001-9446-0136","position":10,"is_corresponding":false},{"id":826308,"name":"Jakub Pawełczyk","orcid":"0000-0001-6863-8594","position":11,"is_corresponding":false},{"id":826309,"name":"Jarosław Dziadek","orcid":"0000-0003-1427-884X","position":12,"is_corresponding":false},{"id":826310,"name":"Jarosław Dastych","orcid":"0000-0001-9102-2261","position":13,"is_corresponding":false},{"id":826299,"name":"Michał Różański","orcid":"0000-0002-7058-836X","position":0,"is_corresponding":true}],"reference_count":34,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-18T23:54:18.470602Z","pmid":null,"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":[]}