Table 2 Primers sequences, cycling conditions and annealing temperature for msp1 and msp2 PCR

msp1

Primary

M1-OF:

CTAGAAGCTTTAGAAGATGCAGTATTG

Initial denaturation: 95 °C for 5 min; PCR: 25 cycles of 94 °C for 1 min, 61 °C for 45 s, 72 °C for 1.5 min; final elongation:72 °C for 5 min

61 °C

M1-OR:

CTTAAATAGTATTCTAATTCAAGTGGATCA

Secondary

M1- KF:

AATGAAGAAGAAATTACTACAAAAGGTGC

Initial denaturation: 95 °C for 5 min; PCR: 30 cycles of 94 °C for 45 s, allele-specific °C for 30s, 72 °C for 1 min; final elongation: 72 °C for 5 min

K1(61 °C)

3D7

M1-KR:

GCTTGCATCAGCTGGAGGGCTTGCACCAGA

M1-MF:

AAATGAAGGAACAAGTGGAACAGCTGTTAC

MAD20 (61 °C)

HB3

M1-MR:

ATCTGAAGGATTTGTACGTCTTGAATTACC

M1-RF:

TAAAGGATGGAGCAAATACTCAAGTTGTTG

R033 (63 °C)

R033

M1-RR:

CATCTGAAGGATTTGCAGCACCTGGAGATC

msp2

Primary

MSP2-1:

ATGAAGGTAATTAAAACATTGTCTATTATA

Initial denaturation: 94 °C for 5 min; PCR: 40 cycles of 94 °C for 1.5 min, 55 °C for 45 s, 72 °C for 1.5 min; final elongation: 72 °C for 10 min

(55 °C)

MSP2-4:

ATATGGCAAAAGATAAAACAAGTGTTGCTG

Secondary

A1:

GCAGAAAGTAAGCCTTCTACTGGTGCT

Initial denaturation: 94 °C for 2 min; PCR: 30 cycles of 94 °C for 1.5 min, 55 °C for 45 s, 72 °C for 1.5 min; final elongation: 72 °C for 10 min

IC/3D7 (55 °C)

3D7

3D7 A2:

GATTTGTTTCGGCATTATTATGA

FC27 B1:

GCAAATGAAGGTTCTAATACTAATAG

FC27 (55 °C)

HB3

FC27 B2:

GCTTTGGGTCCTTCTTCAGTTGATTC