Development of an Integrated Disposable Device for SARS-CoV-2 Nucleic Acid Extraction and Detection

Objective To develop a highly sensitive and rapid nucleic acid detection method for the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).Methods We designed, developed, and manufactured an integrated disposable device for SARS-CoV-2nucleic acid extraction and detection. The precision of the liquid transfer and temperature control was tested. A comparison between our device and a commercial kit for SARS-Cov-2 nucleic acid extraction was performed using real-time fluorescence reverse transcription polymerase chain reaction(RT-PCR).The entire process, from SARS-CoV-2 nucleic acid extraction to amplification, was evaluated.Results The precision of the syringe transfer volume was 19.2 ± 1.9 μL(set value was 20), 32.2 ± 1.6(set value was 30), and 57.2 ± 3.5(set value was 60). Temperature control in the amplification tube was measured at 60.0 ± 0.0 ℃(set value was 60) and 95.1 ± 0.2 ℃(set value was 95) respectively. SARS-Cov-2 nucleic acid extraction yield through the device was 7.10 × 10～6 copies/mL, while a commercial kit yielded 2.98 × 10～6 copies/mL. The mean time to complete the entire assay, from SARS-CoV-2 nucleic acid extraction to amplification detection, was 36 min and 45 s. The detection limit for SARS-CoV-2 nucleic acid was 250 copies/mL.Conclusion The integrated disposable devices may be used for SARS-CoV-2 Point-of-Care test(POCT).