Table 1.
Comparison of different RT-LAMP enabled COVID-19 detection.
| Method | Enzymes/Master Mixes | LAMP primers | LAMP primer targeting gene (N, S, ORF, RdRp, etc.) | Sensitivity | Reference |
|---|---|---|---|---|---|
| RT-LAMP | Bst 2.0 DNA Polymerase WarmStart Reverse Transcriptase |
F3: TCCAGATGAGGATGAAGAAGA B3: AGTCTGAACAACTGGTGTAAG FIP: AGAGCAGCAGAAGTGGCACAGGTGATTGTGAAGAAGAAGAG BIP: TCAACCTGAAGAAGAGCAAGAACTGATTGTCCTCACTGCC LF: CTCATATTGAGTTGATGGCTCA LB: ACAAACTGTTGGTCAACAAGAC |
N gene (speculated from the nucleotide position in the paper) | 1.02 fg/reaction | 7 |
| iLACO | Super- Script® III Reverse Transcriptase First-Strand Synthesis System WarmStart Colormetric LAMP 2X Master Mix |
F3: CCACTAGAGGAGCTACTGTA B3: TGACAAGCTACAACACGT FIP: AGGTGAGGGTTTTCTACATCACTATATTGGAACAAGCAAATTCTATGG BIP: ATGGGTTGGGATTATCCTAAATGTGTGCGAGCAAGAACAAGTG LF: CAGTTTTTAACATGTTGTGCCAACC LB: TAGAGCCATGCCTAACATGCT |
ORF1ab gene | 10 copies/µL | 13, 14 |
| One-pot RT-LAMP | Bst 3.0 DNA/RNA Polymerase |
F3: GCCAAAAGGCTTCTACGCA B3: TTTGGCCTTGTTGTTGTTGG FIP: TCCCCTACTGCTGCCTGGAGTTTTCGGCAGTCAAGCCTCTTC BIP: TCCTGCTAGAATGGCTGGCAATTTTTTTTGCTCTCAAGCTGGTTCA LF: CGACTACGTGATGAGGAACGA LB: GCGGTGATGCTGCTCT |
N gene | 6 copies/μL | 15 |
| Mismatch-tolerant RT-LAMP | WarmStart Reverse Transcriptase Q5® High-Fidelity DNA Polymerase |
F3: TTTGCATTACAGTAGGGATGA B3: TGACAAATATCACTACATCTTTTGG FIP: GCATCGACCACAGAGATAGACACTCAAATGAATCTTAAGTATGCCA BIP: ATAGCCGCCACTAGAGGAGCGCAAATTCTATGGTGGTTGG LF: TAGTGCAAAGAATAGAGCTCGCAC LB: TACTGTAGTAATTGGAACAA |
RdRP in ORF1ab gene | 30 copies/reaction (40 min) 3 copies/reaction (50 min) |
16 |
| LAMP-Seq | Bst 3.0 DNA Polymerase NEBNext 2x Master Mix |
F3: TGGCTACTACCGAAGAGCT B3: TGCAGCATTGTTAGCAGGAT FIP: TCTGGCCCAGTTCCTAGGTAGTNNNNNNNNNNCCAGACGAATTCGTGGTGG BIP: AGACGGCATCATATGGGTTGCACGGGTGCCAATGTGATCT LF: GGACTGAGATCTTTCATTTTACCGT LB: ACTGAGGGAGCCTTGAATACA |
N gene | N/A | 5 |
| RT-LAMP plus sample inactivation (A1) | WarmStart LAMP Kit |
F3: CGGTGGACAAATTGTCAC B3: CTTCTCTGGATTTAACACACTT FIP: TCAGCACACAAAGCCAAAAATTTATCTGTGCAAAGGAAATTAAGGAG BIP: TATTGGTGGAGCTAAACTTAAAGCCCTGTACAATCCCTTTGAGTG LF: TTACAAGCTTAAAGAATGTCTGAACACT LB: TTGAATTTAGGTGAAACATTTGTCACG |
ORF1ab gene | 50 copies/µL | 17 |
| RT-LAMP plus sample inactivation and purification (A1e) | WarmStart LAMP Kit |
F3: Same as A1 B3: Same as A1 FIP: TCAGCACACAAAGCCAAAAATTTATTTTTCTGTGCAAAGGAAATTAAGGAG BIP: TATTGGTGGAGCTAAACTTAAAGCCTTTTCTGTACAATCCCTTTGAGTG LF: Same as A1 LB: Same as A1 |
ORF1ab gene | 1 copy/µL | 17 |
| Penn-RAMP | Isothermal Master Mix (ISO-001, OptiGene) |
F3: TGCTTCAGTCAGCTGATG B3: TTAAATTGTCATCTTCGTCCTT FIP: TCAGTACTAGTGCCTGTGCCCACAATCGTTTTTAAACGGGT BIP: TCGTATACAGGGCTTTTGACATCTATCTTGGAAGCGACAACAA LF: CTGCACTTACACCGCAA LB: GTAGCTGGTTTTGCTAAATTCC |
ORF1ab gene | 7 copies/reaction | 6 |
| RT-LAMP + CRISPR-Cas12 | Bst 2.0 DNA Polymerase WarmStart Reverse Transcriptase |
F3: AACACAAGCTTTCGGCAG B3: GAAATTTGGATCTTTGTCATCC FIP: CGCATTGGCATGGAAGTCACTTTGATGGCACCTGTGTAG BIP: TGCGGCCAATGTTTGTAATCAGCCAAGGAAATTTTGGGGAC LF: TTCCTTGTCTGATTAGTTC LB: ACCTTCGGGAACGTGGTT |
N gene | 10 copies/ μL | 2 |
| STOPCovid | Bst 2.0 WarmStart DNA Polymerase WarmStart Reverse Transcriptase |
F3: GCTGCTGAGGCTTCTAAG B3: GCGTCAATATGCTTATTCAGC FIP: GCGGCCAATGTTTGTAATCAGTAGACGTGGTCCAGAACAA BIP: TCAGCGTTCTTCGGAATGTCGCTGTGTAGGTCAACCACG LF: CCTTGTCTGATTAGTTCCTGGT LB: TGGCATGGAAGTCACACC |
N gene | 100 copies/reaction | 18 |
| Rapid RT-LAMP (N1) | WarmStart Colormetric LAMP 2x Master Mix (DNA & RNA) |
F3: TGGACCCCAAAATCAGCG B3: GCCTTGTCCTCGAGGGAAT FIP: CCACTGCGTTCTCCATTCTGGTAAATGCACCCCGCATTACG BIP: CGCGATCAAAACAACGTCGGCCCTTGCCATGTTGAGTGAGA LF: TGAATCTGAGGGTCCACCAAA LB: GGTTTACCCAATAATACTGCGTCTT |
N gene | 2 copies/reaction (20 min) | 8 |
| Rapid RT-LAMP (N15) | WarmStart Colormetric LAMP 2x Master Mix (DNA & RNA) |
F3: AGATCACATTGGCACCCG B3: CCATTGCCAGCCATTCTAGC FIP: TGCTCCCTTCTGCGTAGAAGCCAATGCTGCAATCGTGCTAC BIP: GGCGGCAGTCAAGCCTCTTCCCTACTGCTGCCTGGAGTT LF: GCAATGTTGTTCCTTGAGGAAGTT LB: GTTCCTCATCACGTAGTCGCAACA |
N gene | 2 copies/reaction (30 min) | 8 |
| Rapid RT-LAMP (S17) | WarmStart Colormetric LAMP 2x Master Mix (DNA & RNA) |
F3: TCTTTCACACGTGGTGTT B3: GTACCAAAAATCCAGCCTC FIP: CATGGAACCAAGTAACATTGGAAAACCTGACAAAGTTTTCAGATCC BIP: CTCTGGGACCAATGGTACTAAGAGGACTTCTCAGTGGAAGCA LF: GAAAGGTAAGAACAAGTCCTGAGT LB: CTGTCCTACCATTTAATGATGGTGT |
S gene | 2 copies/reaction (30 min) | 8 |
| Rapid RT-LAMP (O117) | WarmStart Colormetric LAMP 2x Master Mix (DNA & RNA) |
F3: CCCCAAAATGCTGTTGTT B3: TAGCACGTGGAACCCAAT FIP: GGTTTTCAAGCCAGATTCATTATGGATGTCACAATTCAGAAGTAGGA BIP: TCTTCGTAAGGGTGGTCGCAGCACACTTGTTATGGCAAC LF: TCGGCAAGACTATGCTCAGG LB: TTGCCTTTGGAGGCTGTGT |
ORF1ab gene | 20 copies/reaction (30 min) | 8 |