Validation and implementation of a direct RT-qPCR method for rapid screening of SARS-CoV-2 infection by using non-invasive saliva samples
Validation and implementation of a direct RT-qPCR method for rapid screening of SARS-CoV-2 infection by using non-invasive saliva samples
Abstract BackgroundThere is an urgent need to curb COVID-19 pandemic through early identification of asymptomatic but infectious cases. We aimed to validate and implement an optimised screening method for detection of SARS-CoV-2 RNA combining use of self-collected raw saliva samples, single-step heat-treated virus inactivation and RNA extraction, and direct RT-qPCR. Methods and findingsThe study was conducted in Sant Joan de Déu University Hospital (Barcelona, Spain), including: i) analytical validation against standard RT-qPCR in saliva samples; ii) diagnostic validation against standard RT-qPCR using paired saliva-nasopharyngeal samples obtained from asymptomatic teenagers and young and older adults in a youth sports academy; and iii) high throughput pilot screening of asymptomatic health workers and other staff in the study site.The proposed method had comparable analytical performance to standard RT-qPCR in saliva. Diagnostic validation included saliva samples self-collected with supervision by 173 participants during 9-12 weeks and nasopharyngeal samples collected from them. At baseline, all participants (100.0%) were negative for SARS-CoV-2 in both paired saliva-nasopharyngeal samples. In the following weeks, standard RT-qPCR yielded 23 positive results in nasopharyngeal samples whereas paired saliva specimens yielded 22 (95.7%) positive and one inconclusive result.A total of 2,709 participants engaged in the pilot screening, with high rate of participation (83.4% among health workers). Only 17 (0.6%) of saliva samples self-collected by participants in an unsupervised manner were invalid. Saliva was positive in 24 (0.9%) out of 2,692 valid specimens and inconclusive in 27 (1.0%). All 24 saliva-positive participants and 4 with saliva inconclusive results were positive by standard RT-qPCR in nasopharyngeal samples. The pilot showed potential for rapid analytical workflow (up to 384 batched samples can be processed in <2 hours). ConclusionDirect RT-qPCR on self-collected raw saliva is a simple, rapid, and accurate method with potential to be scaled up for enhanced SARS-CoV-2 community-wide screening.
Bassat Quique、Mu?oz-Almagro Carmen、Torrents Francesc、Claverol Joana、Garcia-Garcia Juan Jose、Brotons Pedro、Rodas Gil、Gratacos Eduard、Fumado Vicky、Launes Cristian、Perez-Arg¨1ello Amaresh、Saucedo Jesica、Jordan Iolanda
Consorcio de Investigaci¨?n Biom¨|dica en Red de Epidemiolog¨aa y Salud P¨2blica (CIBERESP)||Hospital Sant Joan de Deu, Universitat de Barcelona, Esplugues de Llobregat||Centro de Investiga??o em Sa¨2de de Manhi?a (CISM)||ISGlobal, Hospital Cl¨anic - Universitat de Barcelona||ICREA, Pg. Llu¨as Companys 23, 08010 BarcelonaInstitut de Recerca Sant Joan de Deu, Esplugues de Llobregat||Department of Medicine, Universitat Internacional de Catalunya||Consorcio de Investigaci¨?n Biom¨|dica en Red de Epidemiolog¨aa y Salud P¨2blica (CIBERESP)||Hospital Sant Joan de Deu, Universitat de Barcelona, Esplugues de LlobregatHospital Sant Joan de Deu, Universitat de Barcelona, Esplugues de LlobregatFundaci¨? Sant Joan de Deu, Esplugues de LlobregatInstitut de Recerca Sant Joan de Deu, Esplugues de Llobregat||Consorcio de Investigaci¨?n Biom¨|dica en Red de Epidemiolog¨aa y Salud P¨2blica (CIBERESP)||Hospital Sant Joan de Deu, Universitat de Barcelona, Esplugues de LlobregatInstitut de Recerca Sant Joan de Deu, Esplugues de Llobregat||Department of Medicine, Universitat Internacional de Catalunya||Consorcio de Investigaci¨?n Biom¨|dica en Red de Epidemiolog¨aa y Salud P¨2blica (CIBERESP)Medical Department, Futbol Club Barcelona, FIFA Medical Center of Excelence||Medicine Sport Unit, Hospital Clinic and Hospital Sant Joan de DeuBarcelona Center of Maternal-Fetal Medicine and Neonatology (Hospital Clinic and Hospital Sant Joan de Deu), IDIBAPS, University of Barcelona||Centre for Biomedical Research on Rare Diseases (CIBERER)Institut de Recerca Sant Joan de Deu, Esplugues de Llobregat||Hospital Sant Joan de Deu, Universitat de Barcelona, Esplugues de LlobregatInstitut de Recerca Sant Joan de Deu, Esplugues de Llobregat||Consorcio de Investigaci¨?n Biom¨|dica en Red de Epidemiolog¨aa y Salud P¨2blica (CIBERESP)||Hospital Sant Joan de Deu, Universitat de Barcelona, Esplugues de LlobregatInstitut de Recerca Sant Joan de Deu, Esplugues de LlobregatHospital Sant Joan de Deu, Universitat de Barcelona, Esplugues de LlobregatInstitut de Recerca Sant Joan de Deu, Esplugues de Llobregat||Consorcio de Investigaci¨?n Biom¨|dica en Red de Epidemiolog¨aa y Salud P¨2blica (CIBERESP)||Hospital Sant Joan de Deu, Universitat de Barcelona, Esplugues de Llobregat
医学研究方法预防医学基础医学
Bassat Quique,Mu?oz-Almagro Carmen,Torrents Francesc,Claverol Joana,Garcia-Garcia Juan Jose,Brotons Pedro,Rodas Gil,Gratacos Eduard,Fumado Vicky,Launes Cristian,Perez-Arg¨1ello Amaresh,Saucedo Jesica,Jordan Iolanda.Validation and implementation of a direct RT-qPCR method for rapid screening of SARS-CoV-2 infection by using non-invasive saliva samples[EB/OL].(2025-03-28)[2025-04-26].https://www.medrxiv.org/content/10.1101/2020.11.19.20234245.点此复制
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