tetano
Editor, Senior Moderator
Viruses
. 2021 Sep 19;13(9):1875.
doi: 10.3390/v13091875.
A Reverse Transcription Recombinase-Aided Amplification Method for Rapid and Point-of-Care Detection of SARS-CoV-2, including Variants
Fengyun Li[SUP] 1 [/SUP], Ping He[SUP] 2 3 [/SUP], Dongyan Xiong[SUP] 2 3 [/SUP], Yakun Lou[SUP] 4 [/SUP], Qiaosheng Pu[SUP] 1 [/SUP], Haixia Zhang[SUP] 1 [/SUP], Huige Zhang[SUP] 1 [/SUP], Junping Yu[SUP] 2 3 [/SUP]
Affiliations
Abstract
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.
Keywords: RT-RAA; SARS-CoV-2; VOC; variant.
. 2021 Sep 19;13(9):1875.
doi: 10.3390/v13091875.
A Reverse Transcription Recombinase-Aided Amplification Method for Rapid and Point-of-Care Detection of SARS-CoV-2, including Variants
Fengyun Li[SUP] 1 [/SUP], Ping He[SUP] 2 3 [/SUP], Dongyan Xiong[SUP] 2 3 [/SUP], Yakun Lou[SUP] 4 [/SUP], Qiaosheng Pu[SUP] 1 [/SUP], Haixia Zhang[SUP] 1 [/SUP], Huige Zhang[SUP] 1 [/SUP], Junping Yu[SUP] 2 3 [/SUP]
Affiliations
- PMID: 34578456
- DOI: 10.3390/v13091875
Abstract
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.
Keywords: RT-RAA; SARS-CoV-2; VOC; variant.