Table 1.
Primer and probe sequences for the real-time PCR-based detection of Neisseria gonorrhoeae and antimicrobial resistance determinants.
| Assay/target | Primers and probes | Probe sequence 5′ – 3′ |
|---|---|---|
| *PorA1, * | pap-F | CAGCATTCAATTTGTTCCGAGTC |
| pap-R | GAACTGGTTTCATCTGATTACTTTCCA | |
| pap-TM | FAM-CGCCTATACGCCTGCTACTTTCACGC-BHQ1 | |
| *Opa2, * | NGopa-F | TTGAAACACCGCCCGGAA |
| NGopa-R | TTTCGGCTCCTTATTCGGTTTAA | |
| NGopa | TET-CCGATATAATCCGTCCTTCAACATCAG-BHQ1 | |
| GyrA3. & | GyrA-F | TTGCGCCATACGGACGAT |
| GyrA-R | GCGACGTCATCGGTAAATACCA | |
| GyrA91–95 | FAM-TGTCGTAAACTGCGGAA-BHQ-1 | |
| ParC3, & | ParC-F | TGAGCCATGCGCACCAT |
| ParC-R | GGCGAGATTTTGGGTAAATACCA | |
| ParC86–88 | TET-CGGAACTGTCGCCGT-BHQ-1 | |
| PPNG4 | PPNG-F | AGCTGTTCGTTTTTTACTACCAATCA |
| PPNG-R | TGATTTAGTCGTTGAGGTTGAACAA | |
| PPNG-TM | TET-AATTTAAAGAGTGAATAGTACGCCCACGCTTGA-BHQ1 | |
| PBP-25 | NG89-F | GTTGGATGCCCGTACTGGG |
| NG89-R | ACCGATTTTGTAAGGCAGGG | |
| NG89-TM | FAM-CGGCAAAGTGGATGCAACCGA-BHQ-1 |
TM = TaqMan; FAM = Carboxyfluorescein; TET =Tetrachlorofluorescein, IBHQ-1 = Iowa Black Hole Quencher-1; PBP-2 = penicillin-binding protein 2.
1
Assay adapted from Whiley and Sloots 2005.17
2
Assay adapted from Tabrizi et al, 2005.18
3
Assay adapted from Giles et al, 2004.19
4
Assay adapted from Goire et al, 2011.20
5
Assay adapted from Ochiai et al, 2008.21
*
The porA and opa assays were carried out as a multiplex assay.
&
The gyrA and parC assays were carried out as a multiplex assay.