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

Jing Ma, Yan Zhe Hao, Mei Ling Hou, Xiao Shan Zhang, Jing Duan Liu, Hao di Meng, Jiang Bo Chang, Xue Jun Ma, Ji Hua Liu, Qing Jie Ying, Xian Hua Wang, Hong Xia Li, Yu Xi Cao, Xiao Guang Zhang
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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 (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 °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 × 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).

开发用于 SARS-CoV-2 核酸提取和检测的一次性集成装置。
目的开发一种高灵敏、快速的严重急性呼吸系统综合征冠状病毒 2(SARS-CoV-2)核酸检测方法:方法:我们设计、开发并制造了一种用于 SARS-CoV-2 核酸提取和检测的一次性集成装置。测试了液体传输和温度控制的精确性。使用实时荧光反转录聚合酶链反应(RT-PCR)对我们的设备和用于 SARS-Cov-2 核酸提取的商业试剂盒进行了比较。对从 SARS-CoV-2 核酸提取到扩增的整个过程进行了评估:结果:注射器转移量的精确度分别为 19.2 ± 1.9 μL(设定值为 20)、32.2 ± 1.6(设定值为 30)和 57.2 ± 3.5(设定值为 60)。扩增管的温度控制分别为 60.0 ± 0.0 ℃(设定值为 60)和 95.1 ± 0.2 ℃(设定值为 95)。该装置的 SARS-Cov-2 核酸提取率为 7.10 × 10 6 拷贝/毫升,而商业试剂盒的提取率为 2.98 × 10 6 拷贝/毫升。从 SARS-CoV-2 核酸提取到扩增检测,完成整个检测的平均时间为 36 分钟 45 秒:结论:一次性集成装置可用于 SARS-CoV-2 护理点检测(POCT)。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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