4.7 Article

Isothermal gene amplification coupled MALDI-TOF MS for SARS-CoV-2 detection

Journal

TALANTA
Volume 242, Issue -, Pages -

Publisher

ELSEVIER
DOI: 10.1016/j.talanta.2022.123297

Keywords

SARS-CoV-2; Rolling circle amplification; MALDI-TOF MS; Gene detection; Virus analysis

Funding

  1. Ministry of Science and Technology of China [2020YFF0426500, 2020YFF0304502]
  2. National Natural Science Foundation of China (NSFC) [22022401, 22074022, 21934001]

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The researchers developed a new method for the rapid and accurate detection of SARS-CoV-2 using a three-way junction induced exponential rolling circle amplification (3WJ-eRCA) combined with MALDI-TOF MS assay. The method showed high specificity in distinguishing SARS-CoV-2 from other coronaviruses and demonstrated potential for large-scale screening of COVID-19.
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 SARSCoV-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 degrees 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.

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