Centers for Disease Control and Prevention, People's Liberation Army Southern Theater Command, Guangzhou, China.
Laboratory of Medical Mycology, Jining No. 1 People's Hospital, Jining, China.
Lab Med. 2023 Mar 7;54(2):182-189. doi: 10.1093/labmed/lmac093.
The outbreak of COVID-19 caused by SARS-CoV-2 has led to a serious worldwide pandemic. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR)-based methods were recommended for routine detection of SARS-CoV-2 RNA. Because the reaction time and analytical sensitivity of qRT-PCR limits the diagnosis of SARS-CoV-2, development of a quick process of SARS-CoV-2 detection technology with high analytical sensitivity remains urgent.
We combined isothermal amplification and fluorescence detection technology to develop a new auto-recombinase polymerase amplification (RPA)-fluorescence platform that could be used in the diagnosis of SARS-CoV-2.
By optimization of primers and probes, the RPA platform could detect SARS-CoV-2 nucleotides within 15 min. The limits of detection and specificity of the auto-RPA-fluorescence platform were 5 copies/µL and 100%, respectively. The accuracy of detection of the auto-RPA-fluorescence platform in the 16 positive samples was 100%.
The RPA platform is a potential technology for the diagnosis of SARS-CoV-2 infection.
由 SARS-CoV-2 引起的 COVID-19 爆发导致了严重的全球大流行。定量逆转录-聚合酶链反应（qRT-PCR）方法被推荐用于 SARS-CoV-2 RNA 的常规检测。由于 qRT-PCR 的反应时间和分析灵敏度限制了对 SARS-CoV-2 的诊断，因此开发具有高分析灵敏度的 SARS-CoV-2 检测技术的快速过程仍然迫在眉睫。
我们结合等温扩增和荧光检测技术，开发了一种新的自动重组酶聚合酶扩增（RPA）-荧光平台，可用于 SARS-CoV-2 的诊断。
通过优化引物和探针，RPA 平台可在 15 分钟内检测到 SARS-CoV-2 核苷酸。自动 RPA-荧光平台的检测限和特异性分别为 5 拷贝/μL 和 100%。自动 RPA-荧光平台对 16 个阳性样本的检测准确率为 100%。
RPA 平台是 SARS-CoV-2 感染诊断的一种有潜力的技术。
