Gene | PCR round | Primer name | Sequence (5'–3') | Cycling conditions | Positive control |
---|---|---|---|---|---|
msp1 | Primary | N1-Fw | GCAGTATTGACAGGTTATGG | Initial denaturation: 94 °C for 5 min; PCR: 30 cycles of 94 °C for 30 s, 45 °C for 45 s, 70 °C for 1.5 min:; final elongation: 70 °C for 10 min | 3D7, K1, HB3 and Dd2 |
N1-Re | GATTGAAAGGTATTTGAC | ||||
Nested | K1- Fw | ROX/AATGAAGAAGAAATTACTACAAAAGGTGC | Initial denaturation: 94 °C for 5 min; PCR: 30 cycles of 94 °C for 30 s, 52 °C for 45 s, 70 °C for 1.5 min; final elongation: 70 °C for 10 min | 3D7 | |
K1-Re | GTGTCTTGCTTGCATCAGCTGGAGGGCTTGCACCAG | ||||
MAD-Fw | 6FAM/AAATGAAGGAACAAGTGGAACAGCTGTTAC | Dd2 | |||
MAD-Re | GTGTCTTATCTGAAGGATTTGTACGTCTTGAATTACC | ||||
RO33-Fw | HEX/TAAAGGATGGAGCAAATACTCAAGTTGTTG | 7G8 | |||
RO33-Re | GTGTCTTCAAGTAATTTTGAACTCTATGTTTTAAATC | ||||
msp2 | Primary | S2-Fw | GAAGGTAATTAAAACATTGTC | Initial denaturation: 94 °C for 5 min; PCR: 30 cycles of 94 °C for 30 s, 45 °C for 45 s, 70 °C for 1.5 min:; final elongation:70 °C for 10 min | 3D7, K1 and HB3 |
S2-Re | GAGGGATGTTGCTGCTGCTCCACAG | ||||
Nested | Fc 27- Fw | 6FAM/GCATTGCCAGAACTTGAA | Initial denaturation: 94 °C for 5 min; PCR: 30 cycles of 94 °C for 30 s, 52 °C for 45 s, 70 °C for 1.5 min; final elongation: 70 °C for 10 min | Dd2 | |
3D7-fw | HEX/CTGAAGAGGTACTGGTAGA | 3D7 | |||
Stail Re | GTGTCTTGCTTATAATATGAGTATAAGGAGAA | Â |