Biomed Environ Sci.
2024 Jun 20
Jing Ma 1, Yan Zhe Hao 1, Mei Ling Hou 1, Xiao Shan Zhang 1, Jing Duan Liu 2, Hao di Meng 2, Jiang Bo Chang 3, Xue Jun Ma 1, Ji Hua Liu 2, Qing Jie Ying 4, Xian Hua Wang 3, Hong Xia Li 1, Yu Xi Cao 1, Xiao Guang Zhang 1
Abstract
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-2 nucleic 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 μL (set value was 30), and 57.2 ± 3.5 μL (set value was 60). Temperature control in the amplification tube was measured at 60.0 ± 0.0 °C (set value was 60) and 95.1 ± 0.2 °C (set value was 95) respectively. SARS-CoV-2 nucleic acid extraction yield through the device was 7.10 × 10⁶ copies/mL, while a commercial kit yielded 2.98 × 10⁶ copies/mL. The mean time to complete the entire assay, from SARS-CoV-2 nucleic acid extraction to amplification detection, was 36 min 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).
Tags:
An integrated disposable device; Nucleic acid detection; SARS-Cov-2