• Haya Al-Saud, Khaldoun Al-Romaih, Razan Bakheet, Lina Mahmoud, Najla Al-Harbi, Ibtihaj Alshareef, Sara Bin Judia, Layla Aharbi, Abdulaziz Alzayed, Amjad Jabaan, Hani Alhadrami, Ahmed Albarrag, Essam I Azhar, and Maha Ahmad Al-Mozaini.
• From the Saudi Human Genome Project, King Abdulaziz City for Science and Technology, Riyadh, Saudi Arabia.
• Ann Saudi Med. 2020 Sep 1; 40 (5): 373-381.

BackgroundThe 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.ObjectivesReport 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.Materials And MethodsInitially, 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).ResultsAutomated 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.ConclusionsWe 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.LimitationsSmall patient sample size.Conflict Of InterestNone.

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