Figure 3.

Optimization reaction condition of our assay. (A, B) LFD readout of amplicons of ORF1ab gene and N gene in different temperatures ranging from 37°C to 42°C for 30 min using RT-MIRA. (C) LFD readout of amplicons of ORF1ab gene and N gene in different concentrations of primers and probes at 38°C for 30 min using RT-MIRA. (D) LFD readout of amplicons of ORF1ab gene and N gene in twofold diluted volumes of RT-MIRA at 38°C for 30 min. SARS-CoV-2 national reference at 5 × 10⁴ copies/ml were used as template in above experiments. (E, F) LFD readout of RT-MIRA amplicons in different reaction times at 5 × 10⁴ copies or 5 × 10¹ copies/ml.