The worldwide pandemic caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and its emergence of variants needs rapid and point-of-care testing methods for a broad diagnosis. The regular RT-qPCR is time-consuming and limited in central laboratories, so a broad and large-scale screening requirement calls for rapid and in situ methods. In this regard, a reverse transcription recombinase-aided amplification (RT-RAA) is proposed here for the rapid and point-of-care detection of SARS-CoV-2. A set of highly conserved primers and probes targeting more than 98% of SARS-CoV-2 strains, including currently circulating variants (four variants of concerns (VOCs) and three variants of interest (VOIs)), was used in this study. With the preferred primers, the RT-RAA assay showed a 100% specificity to SARS-CoV-2 from eight other respiratory RNA viruses. Moreover, the assay here is of a high sensitivity and 0.48 copies/μL can be detected within 25 min at a constant temperature (42 °C), which can be realized on portable equipment. Furthermore, the RT-RAA assay demonstrated its high agreement for the detection of SARS-CoV-2 in clinical specimens compared with RT-qPCR. The rapid, simple and point-of-care RT-RAA method is expected to be an appealing detection tool to detect SARS-CoV-2, including variants, in clinical diagnostic applications.

中文翻译：

---

一种逆转录重组酶辅助扩增方法，用于快速即时检测 SARS-CoV-2（包括变体）

由严重急性呼吸综合征冠状病毒-2 (SARS-CoV-2) 及其变种的出现引起的全球大流行需要快速的即时检测方法来进行广泛的诊断。常规 RT-qPCR 非常耗时且在中心实验室中受到限制，因此广泛和大规模的筛选要求需要快速和原位方法。在这方面，本文提出了一种逆转录重组酶辅助扩增（RT-RAA），用于快速即时检测 SARS-CoV-2。本研究使用了一组高度保守的引物和探针，针对 98% 以上的 SARS-CoV-2 毒株，包括当前流行的变体（四种关注变体 (VOC) 和三种关注变体 (VOIs)）。使用首选引物，RT-RAA 检测显示对其他 8 种呼吸道 RNA 病毒中的 SARS-CoV-2 具有 100% 的特异性。此外，该检测灵敏度高，在恒温（42℃）下25分钟内即可检测到0.48拷贝/μL，可以在便携式设备上实现。此外，与 RT-qPCR 相比，RT-RAA 检测在临床标本中检测 SARS-CoV-2 方面表现出高度一致性。快速、简单且即时的 RT-RAA 方法有望成为临床诊断应用中检测 SARS-CoV-2（包括变体）的有吸引力的检测工具。