Multiple-centre clinical evaluation of an ultrafast single-tube assay for SARS-CoV-2 RNA

• DOI: 10.1016/j.cmi.2020.05.007
• Publication Date: 2020-05-15T21:22:43Z
• Authors: J. Wang; K. Cai; X. He; X. Shen; J. Wang; J. Liu; J. Xu; F. Qiu; W. Lei; L. Cui; Y. Ge; T. Wu; Y. Zhang; H. Yan; Y. Chen; J. Yu; X. Ma; H. Shi; R. Zhang; X. Li; Y. Gao; P. Niu; W. Tan; G. Wu; Y. Jiang; W. Xu; X. Ma
• Keywords: Microbiology (medical); Adult; Male; 0301 basic medicine; China; Adolescent; Pneumonia, Viral; Real-Time Polymerase Chain Reaction; Sensitivity and Specificity; Betacoronavirus; 03 medical and health sciences; Humans; Child; Pandemics; Aged; Aged, 80 and over; Diagnostic Tests, Routine; SARS-CoV-2; COVID-19; General Medicine; Middle Aged; 3. Good health; Infectious Diseases; RNA, Viral; Original Article; Female; Coronavirus Infections

To evaluate the performance of an ultrafast single-tube nucleic acid isothermal amplification detection assay for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA using clinical samples from multiple centres.A reverse transcription recombinase-aided amplification (RT-RAA) assay for SARS-CoV-2 was conducted within 15 minutes at 39°C with portable instruments after addition of extracted RNA. The clinical performance of RT-RAA assay was evaluated using 947 clinical samples from five institutions in four regions of China; approved commercial fluorescence quantitative real-time PCR (qRT-PCR) kits were used for parallel detection. The sensitivity and specificity of RT-RAA were compared and analysed.The RT-RAA test results of 926 samples were consistent with those of qRT-PCR (330 were positive, 596 negative); 21 results were inconsistent. The sensitivity and specificity of RT-RAA was 97.63% (330/338, 95% confidence interval (CI) 95.21 to 98.90) and 97.87% (596/609, 95% CI 96.28 to 98.81) respectively. The positive and negative predictive values were 96.21% (330/343, 95% CI 93.45 to 97.88) and 98.68% (596/604, 95% CI 97.30 to 99.38) respectively. The total coincidence rate was 97.78% (926/947, 95% CI 96.80 to 98.70), and the kappa was 0.952 (p < 0.05).With comparable sensitivity and specificity to the commercial qRT-PCR kits, RT-RAA assay for SARS-CoV-2 exhibited the distinctive advantages of simplicity and rapidity in terms of operation and turnaround time.