Table 1.
Summary of MPXV diagnostic methods.
| Sr. No | Assay Name | Target Gene | Primers’ Sequences | Probes’ Sequences | Detection Limit | Real-Sample Analysis | References |
|---|---|---|---|---|---|---|---|
| 1 | HA-PCR | HA gene | Forward: 5′-CTGATAATGTAGAAG AC -3′ Reverse: 5′-TTGTATTTACGTGGGTG-3′ |
NA | Not reported | Yes | [52] |
| 2 | ATI-PCR | ATI-gene | Forward: 5′-AATACAAGGAGGATCT-3′ Reverse: 5′-CTTAACTTTTTCTTTTTCTTTCTC-3′ |
NA | Not reported | Yes | [53] |
| 3 | MPXV PCR assay | ATI-gene | Forward: 5′-GAGAGAATCTCTTGATAT-3′ Reverse: 5′-ATTCTAGATTGTAATC-3′ |
NA | Not reported | Yes | [55] |
| 4 | Real-time PCR | B6R | Forward: 5′-ATTGGTCATTATTTTTGTCACAGGAACA-3′ Reverse: 5′-AATGGCGTTGACAATTATGGGTG-3′ |
5′-MGB/DarkQuencher-AGAGATTAGAAATA-3′-FAM | ∼10 viral copies (2 fg) | Yes | [56] |
| 5 | Real-time PCR | G2R | Forward: 5′-CACACCGTCTCTTCCACAGA -3′ Reverse: 5′-GATACAGGTTAATTTCCACATCG -3′ |
5′-FAM AACCCGTCGTAACCAGCAATACATTT-3′-BHQ1 | ∼8.2 genome copies (1.7 fg) | Yes | [62] |
| 6 | Real-time PCR | G2R | Forward: 5′-TGTCTACCTGGATACAGAAAGCAA-3′ Reverse: 5′-GGCATCTCCGTTTAATACATTGAT -3′ |
5′-FAM-CCCATATATGCTAAATGTACCGGTACCGGA-3′-BHQ1 | ∼40.4 copies (9.46 fg) | Yes | [62] |
| 7 | Real-time PCR | F3L | Forward: 5′-CTCATTGATTTTTCG CGGGAT A-3′ Reverse: 5′-GACGATACTCCTCCT CGTTGGT-3′ |
5′-6FAM-CATCAGAATCTGTAGGCCGT-MGBNFQ-3′ | 11–55 fg (50–250 copies) | Yes | [74] |
| 8 | Real-time PCR | N3R | Forward: 5′-AACAACCGT CCTACA ATTAAA CAACA-3′ Reverse: 5′-CGCTATCGAACCATT TTTGTAGTCT-3′ |
5′-6FAM-TAT AAC GGC GAA GAA TAT ACT-MGBNFQ-3′ | 11–55 fg (50–250 copies) | Rodents | [74] |
| 9 | Real-time PCR | B7R | Forward: 5′-ACGTGTTAAACAATGGGTGATG-3′ Reverse: 5′-AACATTTCCATGAATCGTAGTCC-3′ |
5′-TAMRA-TGAATGAATGCGATACTGTATGTGTGGG-3′-BHQ2 | 50 copies per reaction | Yes | [75] |
| 10 | C-LAMP | D14L | FIP-C: 5′-TGGGAGCATTGTAACTTATAGTTGCCCTCCTGAACACATGACA-3′ F3-C: 5′-TGGGTGGATTGGACCATT-3′ BIP-C: 5′-ATCCTCGTATCCGTTATGTCTTCCCACCTATTTGCGAATCTGTT-3′ B3-C: 5′-ATGGTATGGAATCCTGAGG-3′ LOOP-F-C: 5′-GATATTCGTTGATTGGTAACTCTGG-3′ LOOP-C-C: 5′-GTTGGATATAGATGGAGGTGATTGG-3′ |
N/A | 102.4 copies per reaction | Yes | [69] |
| 11 | C-LAMP | ATI | FIP-W: 5′-CCGTTACCGTTTTTACAATCGTTAATCAATGCTGATATGGAAAAGAGA-3′ F3-W: 5′-TACAGTTGAACGACTGCG-3′ BIP-W: 5′-ATAGGCTAAAGACTAGAATCAGGGATTCTGATTCATCCTTTGAGAAG-3′ B3-W: 5′-AGTTCAGTTTTATATGCCGAAT-3′ LOOP-F-W: 5′-GATGTCTATCAAGATCCATGATTCT-3′ LOOP-C-W: 5′-TCTTGAACGATCGCTAGAGA-3′ |
N/A | 103 copies per reaction | Yes | [69] |
| 12 | RPA | G2R | Forward: 5′-AATAAACGGAAGAGATATAGCACCACATGCAC-3′ Reverse: 5′-GTGAGATGTAAAGGTATCCGAACCACACG-3′ |
5′-ACAGAAGCCGTAATCTATGTTGTCTATCGQTFCCTCCGGGAACTTA-3′ | 16 DNA molecules/μL | Yes | [73] |