{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,6,4]],"date-time":"2025-06-04T10:32:36Z","timestamp":1749033156657,"version":"3.40.5"},"reference-count":13,"publisher":"King Faisal Specialist Hospital and Research Centre","issue":"5","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Ann Saudi Med"],"published-print":{"date-parts":[[2020,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:sec>\n            <jats:title>BACKGROUND:<\/jats:title>\n            <jats:p>The pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) has prompted a need for mass testing to identify patients with viral infection. The high demand has created a global bottleneck in testing capacity, which prompted us to modify available resources to extract viral RNA and perform reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) to detect SARS-COV-2.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>OBJECTIVES:<\/jats:title>\n            <jats:p>Report on the use of a DNA extraction kit, after modifications, to extract viral RNA that could then be detected using an FDA-approved SARS-COV-2 RT-qPCR assay.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>MATERIALS AND METHODS:<\/jats:title>\n            <jats:p>Initially, automated RNA extraction was performed using a modified DNA kit on samples from control subjects, a bacteriophage, and an RNA virus. We then verified the automated extraction using the modified kit to detect in-lab propagated SARSCOV-2 titrations using an FDA approved commercial kit (S, N, and ORF1b genes) and an in-house primer-probe based assay (E, RdRp2 and RdRp4 genes).<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>RESULTS:<\/jats:title>\n            <jats:p>Automated RNA extraction on serial dilutions SARS-COV-2 achieved successful one-step RT-qPCR detection down to 60 copies using the commercial kit assay and less than 30 copies using the in-house primer-probe assay. Moreover, RT-qPCR detection was successful after automated RNA extraction using this modified protocol on 12 patient samples of SARS-COV-2 collected by nasopharyngeal swabs and stored in viral transport media.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>CONCLUSIONS:<\/jats:title>\n            <jats:p>We demonstrated the capacity of a modified DNA extraction kit for automated viral RNA extraction and detection using a platform that is suitable for mass testing.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>LIMITATIONS:<\/jats:title>\n            <jats:p>Small patient sample size.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>CONFLICT OF INTEREST:<\/jats:title>\n            <jats:p>None.<\/jats:p>\n          <\/jats:sec>","DOI":"10.5144\/0256-4947.2020.373","type":"journal-article","created":{"date-parts":[[2020,9,20]],"date-time":"2020-09-20T13:50:50Z","timestamp":1600609850000},"page":"373-381","source":"Crossref","is-referenced-by-count":12,"title":["Automated SARS-COV-2 RNA extraction from patient nasopharyngeal samples using a modified DNA extraction kit for high throughput testing"],"prefix":"10.5144","volume":"40","author":[{"given":"Haya","family":"Al-Saud","sequence":"first","affiliation":[{"name":"From the Saudi Human Genome Project, King Abdulaziz City for Science and Technology, Riyadh, Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Khaldoun","family":"Al-Romaih","sequence":"additional","affiliation":[{"name":"From the National Centre of Genomic Technology, King Faisal Specialist Hospital and Research Centre, Riyadh, Saudi 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Research Centre, Riyadh, Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sara Bin","family":"Judia","sequence":"additional","affiliation":[{"name":"From the National Centre of Genomic Technology, King Faisal Specialist Hospital and Research Centre, Riyadh, Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Layla","family":"Aharbi","sequence":"additional","affiliation":[{"name":"From the National Centre of Genomic Technology, King Faisal Specialist Hospital and Research Centre, Riyadh, Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Abdulaziz","family":"Alzayed","sequence":"additional","affiliation":[{"name":"From the Saudi Human Genome Project, King Abdulaziz City for Science and Technology, Riyadh, Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Amjad","family":"Jabaan","sequence":"additional","affiliation":[{"name":"From the Saudi Human Genome Project, King Abdulaziz City for Science and Technology, Riyadh, Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hani","family":"Alhadrami","sequence":"additional","affiliation":[{"name":"From the Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ahmed","family":"Albarrag","sequence":"additional","affiliation":[{"name":"From the Center of Disease Control, Saudi Arabia Ministry of Health, Riyadh, Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Essam I.","family":"Azhar","sequence":"additional","affiliation":[{"name":"From the Department of of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3732-1356","authenticated-orcid":true,"given":"Maha Ahmad","family":"Al-Mozaini","sequence":"additional","affiliation":[{"name":"From the Department of Infection and Immunity, King Faisal Specialist Hospital and Research Centre, Riyadh, Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"3134","reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tim.2016.03.003"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2001017"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1093\/clinchem\/hvaa029"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.2807\/1560-7917.ES.2020.25.3.2000045"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.7326\/M20-1301"},{"key":"e_1_3_4_7_2","doi-asserted-by":"crossref","unstructured":"Vogels CB Brito AF Wyllie AL Fauver JR Ott IM Kalinich CC et al. 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