Abstract
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been spreading worldwide for more than a year and has undergone several mutations and evolutions. Due to the lack of effective therapeutics and long-active vaccines, accurate and large-scale screening and early diagnosis of infected individuals are crucial to control the pandemic. Nevertheless, the current widely used RT-qPCR-based methods suffer from complicated temperature control, long processing time and the risk of false-negative results. Herein, we present a three-way junction induced exponential rolling circle amplification (3WJ-eRCA) combined MALDI-TOF MS assay for SARS-CoV-2 detection. The assay can detect simultaneously the target nucleocapsid (N) and open reading frame 1 ab (orf1ab) genes of SARS-CoV-2 in a single test within 30 min, with an isothermal process (55 °C). High specificity to discriminate SARS-CoV-2 from other coronaviruses, like SARS-CoV, MERS-CoV and bat SARS-like coronavirus (bat-SL-CoVZC45), was observed. We have further used the method to detect pseudovirus of SARS-CoV-2 in various matrices, e.g. water, saliva and urine. The results demonstrated a great potential of the method for large scale screening of COVID-19, which is an important part of the pandemic control.
Keywords: Gene detection; MALDI-TOF MS; Rolling circle amplification; SARS-CoV-2; Virus analysis.
【저자키워드】 SARS-CoV-2, Gene detection, MALDI-TOF MS, Rolling circle amplification, Virus analysis., 【초록키워드】 COVID-19, Saliva, coronavirus, pandemic, Mutation, Vaccines, SARS-CoV, risk, MERS, severe acute respiratory syndrome Coronavirus, MERS-CoV, early diagnosis, SARS-CoV-2 detection, specificity, pseudovirus, nucleocapsid, Urine, temperature, false-negative results, false-negative, Rolling circle amplification, Open reading frame, Analysis, isothermal, water, acute respiratory syndrome, Frame, acute respiratory syndrome coronavirus, acute respiratory syndrome coronavirus 2, other coronaviruses, infected individual, infected individuals, SARS-like coronavirus, effective, current, detect, lack, demonstrated, Rolling, 【제목키워드】 amplification,