期刊
VIRUSES-BASEL
卷 13, 期 9, 页码 -出版社
MDPI
DOI: 10.3390/v13091875
关键词
SARS-CoV-2; variant; VOC; RT-RAA
类别
资金
- COVID-19 control by Hubei province [2020YFC0841200]
- Sino-African Joint Research Center, Chinese Academy of Sciences [SAJC201605, 151542KYSB20200010]
- Gansu Provincial COVID-19 Science and Technology Major Project [20YF2FA020]
The study proposes a rapid and on-site RT-RAA method for the detection of SARS-CoV-2, demonstrating high specificity and sensitivity for various strains, including current variants. This method can quickly detect lower virus copies at a constant temperature, with high agreement compared to RT-qPCR.
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/mu L can be detected within 25 min at a constant temperature (42 degrees 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.
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