Table 1.
Reactivities and specificities of RT-LAMP-BART assays detecting RdRp gene and the L452R spike mutation of SARS-CoV-2.
Table 2.
LAMP primer sets and a probe used in this study.
Fig 1.
A. SARS-RT-LAMP-BART assay data derived via real-time monitoring the lights on the tube. Serial 10-fold-diluted synthetic SARS-CoV-2 RNA including the target region of RdRp and N genes (5×104, 5×103, 5×102, 102, 5×10, and 5 RNA copies) were assayed. The real-time monitoring was performed using Real-time LAMP-BART apparatus (PCRuN, Biogal Galed Lab, Israel). The assay was identified the light output peak in accordance with the manufacturer’s protocol. B. The Real-time RT-PCR (TaqMan probe method) data performed using a commercially available kit, One Step Prime Script RT-PCR kit (Takara Bio, Shiga, Japan), and LightCycler480 (Roche Diagnostics, Basel, Switzerland). Its reaction time was 130 min. (A) Serial 10-fold-diluted synthetic SARS-CoV-2 RNA including the target region of RdRp and N genes (5×104, 5×103, 5×102, 5×10, and 5 RNA copies) plus 2 copies were assayed. (B) The relation between the Crossing point (Cp) of each sample and the log of the amount of initial template RNA.
Table 3.
Detection limits and reaction time of the real-time RT-PCR and RT-LAMP-BART assays used to detect synthetic SARS-CoV-2 RNA including the target region of RdRp and N genes and the synthetic RNA-spiked nasopharyngeal and saliva specimens.
Fig 2.
A. L452R-RT-LAMP-BART assay data with PNA and without PNA derived via real-time monitoring the lights on the tube using 3M™ Molecular Detection Instrument (MDS100, 3M, USA). Assayed samples were synthetic S gene RNA of SARS-CoV-2 with L452R (T1355G) (positive control, 5×104 RNA copies), SARS-CoV-2 RNA (wild-type, JPN/AI/1-004, 5×106 RNA copies), and DW. B. L452R-RT-LAMP-BART assay data derived via real-time monitoring the lights on the tube. Serial 10-fold-diluted samples (synthetic S gene RNA of SARS-CoV-2 with L452R (T1355G); 5×104, 5×103, 5×102, 102, 5×10, and 5 RNA copies) were assayed. The real-time monitoring was performed using Real-time LAMP-BART apparatus (MDS100, 3M, USA). The assay was identified the light output peak in accordance with the manufacturer’s protocol.