Table 2.
Primers and probes used for SARS-CoV-2 detection.
| Gene | Primers | Oligonucleotide Sequence (5′→3′) | Label | Working Concentration |
Amplicon Size bp | Used for | References |
|---|---|---|---|---|---|---|---|
| N | 2019-nCov_N-F | TTACAAACATTGGCCGCAAA | None | 20 μM | End-Point PCR | CDC, 2020 [10] | |
| 2019-nCov_N-R | GCGCGACATTCCGAAGAA | None | 20 μM | 66 | RT-qPCR | ||
| 2019-nCov_N-P | FAM-ACAATTTGCCCCCAGCGCTTCAG -BHQ1 | FAM, BHQ-1 | 5 μM | ddPCR | |||
| Spike | 2019-nCov_Spike-F | CGGCCTTACTGTTTTGCCAC | None | 20 μM | 75 bp | Syber Green | Falzone et al., 2020 [12] |
| 2019-nCov_Spike-R | TGTACCCGCTAACAGTGCAG | None | 20 μM | RT-qPCR | |||
| Spike/Orfa3 | 2019-nCov_Spike/Orfa3-F | TGAGCCAGTGCTCAAAGGAG | None | 20 μM | 195 | End-Point PCR | this paper |
| 2019-nCov_Spike/Orfa3-R | CGCCAACAATAAGCCATCCG | None | 20 μM | ||||
| GAPDH | hGAPDH-F | CATGAGAAGTATGACAACAGCC | None | 20 μM | 115 | RT-qPCR | Carpenter et al., 2005 [14] |
| hGAPDH-R | TGAGTCCTTCCACGATACC | None | 20 μM | ddPCR | |||
| hGAPDH-P | FAM- AGCAATGCCTCCTGCACCACCAA -BHQ1 | FAM, BHQ-1 | 5 μM |
The table reports the oligonucleotide sequence for N gene, Spike gene, Spike/Orfa3 and Human Control gene GAPDH, label, working concentration, amplicon size and technique used for each gene amplified. Primers for Spike/Orfa3 were designed and analyzed with the online software: http://www.premierbiosoft.com/netprimer/. F, forward primers; R, reverse primers; P, probe.