Full-in-House Method (FinHM) for SARS-COV-2 Automated Viral RNA Extraction, Followed by in-House ‘Primer-Probe’ Based RT-qPCR Detection; Low Cost Mass Testing

Background: Sever acute respiratory syndrome Coronavirus-2 (SARS-COV-2) spread prompted mass testing. The main method for testing is by any FDA approved kits RNA extraction followed One-Step RT-qPCR based on primer-probe assays. Yet, the high demand these created a global bottleneck in capacity. Methods: We developed Full-In-House Method (FinHM) suitable automated viral using full in-house solutions utilizing MagMaxTM beads an In-House CDC/WHO recommended ‘primer-probe’ assay targeting following genes; E, RdRp2, and RdRp4. FinHM was validated kit that targets S, N, ORF1b genes made Thermo Fisher Scientific (TF). Results: sensitivity specificity of were evaluated serial dilutions in-laboratory propagated SARS-COV-2 with successful detection down to 46 copies both assays (P>0.05). Moreover, 266 clinical samples, which test results replicated (>99% similarity, P>0.05). had low limit (5 templates), significant negative correlation between Ct values titrations as shown Pearson (-0.8, -0.8 -0.7 RdRp2 RdRp4, respectively). Finally, also SARS-COV-2-spiked plasma patient samples. Conclusion: report reliable, reproducible, specific, sensitive low-cost platform from other viruses