key: cord-331807-ooym5eh3 authors: Wu, Tao; Ge, Yiyue; Zhao, Kangchen; Zhu, Xiaojuan; Chen, Yin; Wu, Bin; Zhu, Fengcai; Zhu, Baoli; Cui, Lunbiao title: A reverse-transcription recombinase-aided amplification assay for the rapid detection of N gene of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) date: 2020-07-29 journal: Virology DOI: 10.1016/j.virol.2020.07.006 sha: doc_id: 331807 cord_uid: ooym5eh3 The current outbreak of coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was reported in China firstly. A rapid, highly sensitive, specific, and simple operational method was needed for the detection of SARS-CoV-2. Here, we established a real-time reverse-transcription recombinase-aided amplification assay (RT-RAA) to detect SARS-CoV-2 rapidly. The primers and probe were designed based on the nucleocapsid protein gene (N gene) sequence of SARS-CoV-2. The detection limit was 10 copies per reaction in this assay, which could be conducted within 15 min at a constant temperature (39 °C), without any cross-reactions with other respiratory tract pathogens, such as other coronaviruses. Furthermore, compared with commercial real-time RT-PCR assay, it showed a kappa value of 0.959 (p < 0.001) from 150 clinical specimens. These results indicated that this real-time RT-RAA assay may be a valuable tool for detecting SARS-CoV-2. RT-RAA assays were manually designed based on the sequence of N gene (Table 1) . Nuclease-free water was used as a negative control. The sensitivity of RT-RAA assay for SARS-CoV-2 was determined using a panel of 124 serially diluted recombinant plasmid ranging from 10 1 to 10 5 copies per reaction. The 125 repeatability of the method was evaluated by testing each dilution 8 times (Table 2 ). The minimum detection limit of real-time RAA assay was 10 copies / reaction. As As showed in Table 3, Pathogen Genomics in Public Health Use of a rapid 207 reverse-transcription recombinase aided amplification assay for respiratory syncytial virus detection Epidemiological and clinical characteristics of 99 cases of 2019 novel coronavirus pneumonia in 211 China: a descriptive study Detection of 2019 novel coronavirus (2019-nCoV) by real-time 214 RT-PCR Clinical Features Predicting Mortality 216 Risk in Patients With Viral Pneumonia: The MuLBSTA Score Genomic characterisation and epidemiology of 2019 novel coronavirus: implications for virus 221 origins and receptor binding Assessment of the quantitative real-time polymerase chain reaction using a cDNA standard for 224 human group A rotavirus Notice on novel coronavirus pneumonia diagnosis and treatment plan (trial version fifth 226 Revision) A New Arenavirus in a Cluster of Fatal Transplant-Associated Diseases DNA Detection Using Recombination 231 Proteins 2019 novel coronavirus of pneumonia in Wuhan, 233 China: emerging attack and management strategies A rapid 235 and sensitive recombinase aided amplification assay to detect hepatitis B virus without DNA 236 extraction Multiple-centre clinical evaluation of an ultrafast single-tube assay for SARS-CoV-2 RNA Clinical microbiology and infection : the official publication of the European Society of Clinical 241 Microbiology and Infectious Diseases Far-Eastern Subtype of Tick-borne Encephalitis Virus WHO, 2020. Coronavirus disease 2019 (COVID-19) Situation Report -35 Development of a reverse 248 transcription recombinase-aided amplification assay for the detection of coxsackievirus A10 and 249 coxsackievirus A6 RNA Rapid detection 251 of Salmonella with Recombinase Aided Amplification A Novel 254 Coronavirus from Patients with Pneumonia in China