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Table 1 Primer pairs used to amplify and sequence pfcrt, pfmdr1 and pvmdr1

From: Return of chloroquine-sensitive Plasmodium falciparum parasites and emergence of chloroquine-resistant Plasmodium vivax in Ethiopia

Gene fragment

Primer Name

Primer sequence

PCR cycling conditions

Pfcrt SNPs at

OF P1

5′CCGTTAATAATAAATACACGCG

35 cycles of 94°C for 30 s; 56°C

Codon 72&76

OR P2

5′CGGATGTTACAAAACTATAGTCC

for 30 s; and 62°C for 1 min;

 

NF P3

5′-AGGTTCTTGTCTTGGTAAATTTGC

30 cycles of 94°C for 30 s; 56°C

 

NR P4

5′-CAAAACTATAGTTACCAATTTTG

for 30 s; and 65°C for 1 min;

Pfmdr1 SNPs at

OF P5

5′-AGGTTGAAAAAGAGTTGAAC

30 cycles of 94°C for 30 s; 55°C

Codon 86&184

OR P6

5′-ATGACACCACAAACATAAAT

for 30 s; and 65°C for 1 min;

 

NF P7

5′-ACAAAAAGAGTACCGCTGAAT

30 cycles of 94°C 30 s; 60°C

 

NR P8

5′-AAACGCAAGTAATACATAAAGTC

for 30 s; and 65°C for 1 min;

Pfmdr1 SNPs at

OF P9

5′-GTGTATTTGCTGTAAGAGCT

34 cycles of 94°C for 30 s; 55°C

Codon 1034

OR P10

5′-GACATATTAAATAACATGGGTTC

for 1 min and 72°C for 1.5

1042 and 1246

NF P11

5′ CAGATGATGAAATGTTTAAAGATC

29 cycles of 94°C for 30 s; 60°C

 

NR 12

5′-TAAATAACATGGGTTCTTGACT

for 30 s; and 65°C for 1 min;

pvmdr1at codon

OF P13

5′-GCGAACTCGAATAAGTACTCCCTCTA

45 cycles of 94°C for 5 min,

976 and 1076

OF P14

5′GGCGTAGCTTCCCGTAAATAAA

530C for 1 min and 720C for 1 min

 

NF P15

5′-GGATTGCTGTCAGCACATATTAACA

45 cycles of 94°C for 5 min,

 

NF P16

5′AGAGGGATTTCATAAAGTCATT

650C for 1 min and 720C for 1 min

  1. OF: Outer Forward; OR: Outer Nested; P1-P12: Primers; NF: Nested Forward; NR: Nested Reverse; bp: base pair; pfmdr1: Plasmodium falciparum multi-drug resistance 1; pfcrt: Plasmodium falciparum chloroquine resistance transporter; pvmdr1: Plasmodium. vivax multi-drug resistance 1.