Background The ongoing outbreak of the novel human coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) (also known as 2019-nCoV) has become a global health concern. Rapid and easy-to-use diagnostic techniques are urgently needed to diagnose SARS-CoV-2 infection. Methods We devised a reverse transcription multiple cross-displacement amplification (RT-MCDA) coupled with a nanoparticle-based biosensor assay (RT-MCDA-BS) for rapid, sensitive and specific diagnosis of coronavirus disease 2019 (COVID-19). Two primer sets were designed to target the open reading frame 1a/b and nucleoprotein gene of SARS-CoV-2. A total of 183 clinical samples, including 65 patients with COVID-19 infection and 118 patients with other pathogen infections were used to testify the assay’s feasibility. Diagnosis results were reported visually using the biosensor. Findings The assay designed was performed using a simple instrument which could maintain the reaction in a constant temperature at 64°C for only 35 min. The total COVID-19 RT-MCDA-BS test procedure could be finished within 1 h. The COVID-19 RT-MCDA-BS could detect down to five copies of target sequences. Among 65 clinical samples from the COVID-19 patients, 22 (33.8%) positive results were obtained from faeces, nasal, pharyngeal and anal swabs via COVID-19 RT-MCDA-BS assay, while real-time reverse transcription-PCR assay only detected 20 (30.7%) positive results in these samples. No positive results were obtained from clinical samples with non-COVID-19 infections. Interpretation COVID-19 RT-MCDA-BS was a rapid, reliable, low-cost and easy-to-use assay, which could provide an attractive laboratory tool to diagnose COVID-19 in multiple clinical specimens, especially for field, clinic laboratories and primary care facilities in resource-poor settings. We devised a novel method (COVID-19 RT-MCDA-BS) to diagnose COVID-19. Its rapidity, low cost and ease of use make the method an ideal tool for use in field, primary and clinical laboratories, especially for resource-poor settings. https://bit.ly/3j08CU2
【초록키워드】 COVID-19, coronavirus disease, SARS-CoV-2, Coronavirus disease 2019, coronavirus, primary care, feasibility, SARS-COV-2 infection, 2019-nCoV, Infection, diagnostic, Diagnosis, nasal, severe acute respiratory syndrome Coronavirus, reverse transcription-PCR, Laboratory, biosensor, amplification, clinical samples, Health, infections, pathogen, COVID-19 infection, outbreak, Patient, nanoparticle, Rapid, temperature, reverse transcription, anal swab, respiratory, anal swabs, specimens, COVID-19 patients, diagnose, Open reading frame, Faeces, clinical laboratories, Non-COVID-19, acute respiratory syndrome, Frame, acute respiratory syndrome coronavirus, reaction, positive result, PCR assay, finding, pharyngeal, open reading frame 1, FIVE, human coronavirus, clinical sample, detect, primer set, reported, was performed, were used, maintain, patients with COVID-19, 【제목키워드】 diagnosis of COVID-19,