Simultaneous Detection of SARS-CoV-2 and Six Other Human Coronaviruses
by Multiplex PCR and MALDI-TOF MS
Abstract
The outbreak of severe acute respiratory syndrome coronavirus 2
(SARS-CoV-2) is challenging the health systems worldwide, and large
population testing is a vital step to control this pandemic. Here, we
developed a new method (named HCoV-MS), which combines multiplex PCR
with matrix-assisted laser desorption/ionization-time of flight mass
spectrometry to simultaneously detect and differentiate seven human
coronaviruses (HCoVs). The HCoV-MS method had good specificity and
sensitivity, with a detection limit of 1-5 copies/reaction. To validate
the HCoV-MS method, we tested 151 clinical samples, and the results
showed good concordance with real-time PCR. In addition, 41 D614G
variants were identified, which were consistent with the sequencing
results. This method was also used in EQAE-SARS-COV in 2020, and all the
samples were accurately identified. Taken together, HCoV-MS could be
used as an effective method for large-scale detection. It was also
capable of detecting key single nucleotide polymorphism about variants.