Abstract
Regular, accurate, rapid, and inexpensive self-testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is urgently needed to quell pandemic propagation. The existing at-home nucleic acid testing (NAT) test has high sensitivity and specificity, but it requires users to mail the sample to the central lab, which often takes 3-5 days to obtain the results. On the other hand, rapid antigen tests for the SARS-CoV-2 antigen provide a fast sample to answer the test (15 min). However, the sensitivity of antigen tests is 30 to 40% lower than nucleic acid testing, which could miss a significant portion of infected patients. Here, we developed a fully integrated SARS-CoV-2 reverse transcription loop-mediated isothermal amplification (RT-LAMP) device using a self-collected saliva sample. This platform can automatically handle the complexity and can perform the functions, including (1) virus particles’ thermal lysis preparation, (2) sample dispensing, (3) target sequence RT-LAMP amplification, (4) real-time detection, and (5) result report and communication. With a turnaround time of less than 45 min, our device achieved the limit of detection (LoD) of 5 copies/μL of the saliva sample, which is comparable with the LoD (6 copies/μL) using FDA-approved quantitative real-time polymerase chain reaction (qRT-PCR) assays with the same heat-lysis saliva sample preparation method. With clinical samples, our platform showed a good agreement with the results from the gold-standard RT-PCR method. These results show that our platform can perform self-administrated SARS-CoV-2 nucleic acid testing by laypersons with noninvasive saliva samples. We believe that our self-testing platform will have an ongoing benefit for COVID-19 control and fighting future pandemics.
Keywords: POCT; RT-LAMP; SARS-CoV-2; saliva; self-testing.
【저자키워드】 SARS-CoV-2, Saliva, RT-LAMP, POCT, Self-testing., 【초록키워드】 COVID-19, Saliva, coronavirus, pandemic, communication, qRT-PCR, severe acute respiratory syndrome Coronavirus, RT-PCR, Antigen, polymerase chain reaction, amplification, clinical samples, nucleic acid, RT-LAMP, sensitivity, Rapid antigen tests, nucleic acid testing, rapid antigen test, Sensitivity and specificity, Loop-mediated isothermal amplification, LAMP, limit of detection, POCT, Pandemics, reverse transcription, isothermal amplification, antigen tests, SARS-CoV-2 antigen, Quantitative, platform, noninvasive, Chain Reaction, miss a, acute respiratory syndrome, acute respiratory syndrome coronavirus, infected patients, acute respiratory syndrome coronavirus 2, complexity, sequence, SARS-CoV-2 nucleic acid, functions, lab, benefit, polymerase chain, virus particle, less, comparable, automatically, Regular, the SARS-CoV-2, 【제목키워드】 Device, Mobile,