Primers and probes (concentrations) | 5′ modifications | Sequences | Reaction conditions | References | |
---|---|---|---|---|---|
An. gambiae s.l. mutation | |||||
L1014F-kdr | IPCF (2.5 pmol/µL) AltRev (2.5 pmol/µL) WT (25 pmol/µL) West (8 pmol/µL) | GATAATGTGGATAGATTCCCCGACCATG TGCCGTTGGTGCAGACAAGGATG GGTCCATGTTAATTTGCATTACTTACGAATA CTTGGCCACTGTAGTGATAGGAAATGTT | 1 cycle: 5 min at 95 °C 35 cycles: 30 s at 95 °C, 30 s at 59 °C, 30 s at 72 °C 1 cycle: 5 min at 72 °C | [25] | |
L1014S-kdr | IPCF (2.5 pmol/µL) AltRev (2.5 pmol/µL) WT (5 pmol/µL) East (2.5 pmol/µL) | GATAATGTGGATAGATTCCCCGACCATG TGCCGTTGGTGCAGACAAGGATG GGTCCATGTTAATTTGCATTACTTACGAATA CTTGGCCACTGTAGTGATAGGAAAATC | 1 cycle: 5 min at 95 °C 35 cycles: 30 s at 95 °C, 30 s at 57 °C, 30 s at 72 °C 1 cycle: 5 min at 72 °C | [25] | |
N1575Y | Forward (1 µM) Reverse (1 µM) N (0.5 µM) Y (0.5 µM) | HEX FAM | TGGATCGCTAGAAATGTTCATGACA CGAGGAATTGCCTTTAGAGGTTTCT ATTTTTTTCATTGCATTATAGTAC TTTTTCATTGCATAATAGTAC | 1 cycle: 15 min at 95 °C 40 cycles: 15 s at 94 °C, 1 min at 60 °C | [24] |
G119S-ace1 | Forward (0.8 µM) Reverse (0.8 µM) G (0.2 µM) S (0.2 µM) | HEX FAM | GGCCGTCATGCTGTGGAT GCGGTGCCGGAGTAGA TTCGGCGGCGGCT TTCGGCGGCAGCT | 1 cycle: 10 min at 95 °C 40 cycles: 10 s at 95 °C, 35 s at 60 °C | [25] |
L43F-CYP4J5 | Forward (1 µM) Reverse (1 µM) Probe 1 (0.5 µM) Probe 2 (0.5 µM) | FAM HEX | AGCCTGCGCGTGTGATA CTTCTTCTCCTGTGGTTCGTTTG TTGCCGGAAGGCAGT TTGCCGGAGGGCAGT | 1 cycle: 10 min at 95 °C 40 cycles: 15 s at 92 °C, 1 min at 60 °C | [26] |
CYP6P4-I236M | CYP6P4_I236M_Forward (0.35 µM) CYP6P4_I236M_Reverse (0.35 µM) CYP6P4_I-Wild (0.2 µM) CYP6P4_M-Mutant (0.2 µM) | FAM HEX | AGTTTATGTTTGCGACCACGTT TCCACCGTCTCGCGCACAAC TTC+ATGC+C+G+ATGC TTC+ATGC+C+C+ATGC | 1 cycle: 3 min at 95 °C 20 cycles: 15 s at 95 °C, 30 s at 66 °C 23 cycles: 10 s at 95 °C, 20 s at 58 °C, 11 s at 72 °C | [27] |
Zanzibar-like TE | ZZB_Flank_A (0.35 µM) ZZB_Flank_B (0.35 µM) ZZB_Int_A (0.35 µM) ZZB_Mutant (0.2 µM) ZZB_WT (0.2 µM) | FAM HEX | CAAAATCAATGKCACRGAGC CGCTACAATGAAKGGAAAGTC CATTACATGGCGACCGTACCT A+C+ATTA+CA+CTTTGT+CA+GTA GATG+TT+CTTTK+T+CA+G+TATT | 1 cycle: 3 min at 95 °C 40 cycles: 10 s at 95 °C, 20 s at 58 °C, 11 s at 72 °C | [27] |
CYP6AA1 duplication | AA1_Dup1_ins1 (0.35 µM) AA1_Dup1_outs (0.35 µM) AA1_Dup1_ins2 (0.35 µM) AA1_Dup1_outs (0.2 µM) AA1_Dup1_ins (0.2 µM) AA1_Dup1_junct (0.2 µM) | Cy5 HEX FAM | CAGTGCGGTACGCTCGTTAA GGATCGGTTTACAGCGGACG CATCACCTGTGCTCGCAARTT CCAT+CA+C+CGAA+CG AGAA+CCTGCA+C+CAA A+CA+AT+TAATT+G+CAT+CGG | 1 cycle: 3 min at 95 °C 40 cycles: 10 s at 95 °C, 20 s at 57 °C, 15 s at 72 °C | [27] |
2La inversion | 23A2 (25 pmol/µL) 27A2 (25 pmol/µL) DPCross 52 L+ (25 pmol/µL) | CTCGAAGGGACAGCGAATTA ACACATGCTCCTTGTGAACG GGTATTTCTGGTCACTCTGTTGG | 1 cycle: 2 min at 94 °C 35 cycles: 30 s at 94 °C, 30 s at 60 °C, 45 s at 72 °C 1 cycle: 10 min at 72 °C | [25] | |
An. funestus s.l. mutation | |||||
L119F-GSTe2 | Forward (1 µM) Reverse (1 µM) L119 (0.5 µM) 119 F (0.5 µM) | HEX FAM | AACAATTTTTCATTTCTTATTCTCATTTAC CGACTCGATCTTCGGGAATGTC AGGAGCGTATTCTTTTCTAC AGGAGCGTATTTTTTTCTA | 1 cycle: 10 min at 95 °C 40 cycles: 15 s at 92 °C and 1 min at 60 °C | [28] |
CYP6P9a | Forward (1 µM) Reverse (1 µM) | TCCCGAAATACAGCCTTTCAG ATTGGTGCCATCGCTAGAAG | 1 cycle: 95 °C for 3 min 40 cycles: 94 °C for 30 s, 55 °C for 30 s, 72 °C for 1 min 1 cycle: 72 °C for 10 min | [18] | |
CYP6P9b | Forward (1 µM) Reverse (1 µM) | CCCCCACAGGTGGTAACTATCTGAA TTATCCGTAACTCAATAGCGATG | 1 cycle: 95 °C for 3 min 40 cycles: 94 °C for 30 s, 58 °C for 30 s, 72 °C for 1 min 1 cycle: 72 °C for 10 min | [19] |