Abstract
The current pandemic of COVID-19 caused by SARS-CoV-2 (severe acute respiratory syndrome coronavirus-2) has raised significant public health concerns. Rapid and accurate testing of SARS-CoV-2 is urgently needed for early detection and control of the disease spread. Here, we present an RT-LAMP coupled glass nanopore digital counting method for rapid detection of SARS-CoV-2. We validated and compared two one-pot RT-LAMP assays targeting nucleocapsid (N) and envelop (E) genes. The nucleocapsid assay was adopted due to its quick time to positive and better copy number sensitivity. For qualitative positive/negative classification of a testing sample, we used the glass nanopore to digitally count the RT-LAMP amplicons and benchmarked the event rate with a threshold. Due to its intrinsic single molecule sensitivity, nanopore sensors could capture the amplification dynamics more rapidly (quick time to positive). We validated our RT-LAMP coupled glass nanopore digital counting method for SARS-CoV-2 detection by using both spiked saliva samples and COVID-19 clinical nasopharyngeal swab samples. The results obtained showed excellent agreement with the gold standard RT-PCR assay. With its integration capability, the electronic nanopore digital counting platform has significant potential to provide a rapid, sensitive, and specific point-of-care assay for SARS-CoV-2.
Keywords: COVID-19; Clinical; RT-LAMP; SARS-CoV-2; Solid-state nanopore.
【저자키워드】 COVID-19, SARS-CoV-2, RT-LAMP, clinical, Solid-state nanopore., 【초록키워드】 public health, Saliva, Nanopore, severe acute respiratory syndrome Coronavirus, Spread, Severe acute respiratory syndrome, Nasopharyngeal swab, point-of-care, amplification, RT-LAMP, SARS-CoV-2 detection, sensitivity, nucleocapsid, Early detection, Rapid, gold, respiratory, disease spread, threshold, RT-PCR assay, platform, Coronavirus-2, Amplicon, copy number, capture, acute respiratory syndrome, acute respiratory syndrome coronavirus, gold standard, acute respiratory syndrome coronavirus-2, event rate, saliva samples, positive, pandemic of COVID-19, Genes, intrinsic, caused, the disease, raised, adopted, 【제목키워드】 Nanopore, Rapid, novel coronavirus SARS-CoV-2,