Reverse-Transcription Recombinase-Aided Amplification Assay for Rapid Detection of the 2019 Novel Coronavirus (SARS-CoV-2)

Abstract

A novel coronavirus (SARS-CoV-2) was recently identified in patients with acute respiratory disease and spread quickly worldwide. A specific and rapid diagnostic method is important for early identification. The reverse-transcription recombinase-aided amplification (RT-RAA) assay is a rapid detection method for several pathogens. Assays were performed within 5–15 min as a one-step single tube reaction at 39 °C. In this study, we established two RT-RAA assays for the S and orf1ab gene of SARS-CoV-2 using clinical specimens for validation. The analytical sensitivity of the RT-RAA assay was 10 copies for the S and one copy for the orf1ab gene per reaction. Cross-reactions were not observed with any of the other respiratory pathogens. A 100% agreement between the RT-RAA and real-time PCR assays was accomplished after testing 120 respiratory specimens. These results demonstrate that the proposed RT-RAA assay will be beneficial as it is a faster, more sensitive, and more specific tool for the detection of SARS-CoV-2.

The estimation of diagnostic accuracy of tests for COVID-19: A scoping review

“… 24 Two studies, Wang, Cai, & He et al and Xue et al, evaluated reverse-transcription recombinase-aided amplification (RT-RAA) with Cohen's kappa of 0.952 and 1.0. 26 , 27 Two studies, Perchetti et al and Waggoner et al, evaluated triplex rRT-PCR, reporting overall agreement as 99.2 and 100%. 28 , 29 Li et al evaluated an automatic integrated gene detection system (AIGS) with rSN of 97.2% and rSp 98.5%.…”

“…Regarding evaluation of quality (Supplementary Table 3), the majority of studies did not report how patient samples were selected for evaluation. 21 , 22 , 24 , 25 , 27 , 28 , 29 , 30 , 32 In the study conducted by Bulterys et al, sample selection was a convenience selection of samples with residual volume that had been stored correctly. 31 Most studies did not report symptomatic status of the patient 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 30 , 31 , 32 or patient demographics.…”

A serological aptamer-assisted proximity ligation assay for COVID-19 diagnosis and seeking neutralizing aptamers

“…To this aim, recent advances in molecular biology have produced various nucleic acid detection methods based on the conserved genome sequences of COVID-19, among which real-time quantitative polymerase chain reaction (RT-qPCR) is recommended as a gold standard for COVID-19 diagnostics. [2][3][4] Despite the progress made, RT-qPCR tests have still been reported for false-negative results in some suspected cases with typical clinical characteristics of COVID-19 and identical specic computed tomography (CT) images. [5][6][7] In addition, some mild or asymptomatic cases with initially negative diagnosis were conrmed with COVID-19 infection only aer second, third or multiple repeated RT-qPCR tests.…”

The Potential role of Procyanidin as a Therapeutic Agent against SARS-CoV-2: A Text Mining, Molecular Docking and Molecular Dynamics Simulation Approach

“…Transition between a metastable prefusion state to a stable post-fusion state is triggered by the binding of spike protein with ACE2. Several researchers studied and reviewed the binding mechanism of spike protein with ACE2 receptor and screened small molecule and other inhibitors towards different targets [18][19][20][21][22][23][24][25][26][27][28][29][30]. Also, the receptor-binding domain (RBD) dynamics and development of inhibitor targeting the RBD domain [31] and suggested the potential role of the RBD domain towards the therapeutic development against SARS CoV2.…”