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Volume 28, Number 4—April 2022
Research

Molecular Surveillance for Imported Antimicrobial Resistant Plasmodium falciparum, Ontario, Canada

Ruwandi Kariyawasam, Rachel Lau, Eric Shao, Katherine Tan, Adrienne Showler, Filip Ralevski, Samir N. Patel, and Andrea K. BoggildComments to Author 
Author affiliations: University of Alberta, Edmonton, Alberta, Canada (R. Kariyawasam); Alberta Precision Labs-Public Health, Edmonton (R. Kariyawasam); Public Health Ontario Laboratory, Toronto, Ontario, Canada (R. Lau, F. Ralevski, S.N. Patel); University of Western Ontario, London, Ontario (E. Shao); University of Toronto, Toronto (K. Tan, A. Showler, S.N. Patel, A.K. Boggild); Georgetown University, Washington, DC, USA (A. Showler); Toronto General Hospital, Toronto (A. Showler, A.K. Boggild)

Main Article

Table 1

Primer sets used for pyrosequencing and Sanger sequencing for Plasmodium falciparum resistance testing*

Primer† Sequence, 5′ → 3′
Pf ATPase 6 codon 623
Forward primer CACCAAGGGGTATCAACAAATTC
Biotinylated reverse primer (HPLC purified) CTTGTGCCATGCTCAAATGTATT
Sequence primer
 TCTAATTATACTACAGCTCA
Pf ATPase 6 codon 769
Biotinylated forward primer (HPLC purified) CAAAATATGGGAAAAAGAGCATTA
Reverse primer TCTGGCCGTATTAATATTATCACC
Sequence primer
 ATAAATTAAATCTTGTTCTA
Pf cytb
Forward primer CGTCCTGATAATGCTATCGTAGTAAATAC
Biotinylated reverse primer (HPLC purified) CAGGCATCTAACATGGATATAGCA
Sequence primer
 TGGTACTTTCTACCATT
pfcrt
Biotinylated forward primer (HPLC purified) GGAGGTTCTTGTCTTGGTAAA
Reverse primer CAGGCATCTAACATGGATATAGCA
Sequence primer
 TTTGTTTAAAGTTCTTTTAG
Pf MDR1 codon 86
Forward primer CGTTTAAATGTTTACCTGCACAA
Biotinylated reverse primer (HPLC purified) TTGTCCATCTTGATAAAAAACACT
Sequence primer
 GTGTAATATTAAAGAACATG
Pf MDR1 codon 184
Forward primer AGTTCAGGAATTGGTACGAAATTT
Biotinylated reverse primer (HPLC purified) AACGTGCATTTTTTATTAATGACC
Sequence primer
 CCAGTTCCTTTTTAGGTT
Pf MDR1 codon 1034, 1042
Forward primer GCGGAGTTTTTGCATTTAGTTCA
Biotinylated reverse primer (HPLC purified) TTAAGAAGGATCCAAACCAATAGG
Sequence primer
 GCAGCTTTATGGGGAT
Pf MDR1 codon 1246
Biotinylated forward primer (HPLC purified) TTTTCAAACCAATCTGGATCTG
Reverse primer CGTTTAACATCTTCCAATGTTGC
Sequence primer
 ATTGAAAATAAGTTTCTAAG
Pf dhfr codon 16
Biotinylated forward primer AACAAGTCTGCGACGTTTTCGATA
Reverse primer CCCCTCATTTTTGCTTTCAACC
Sequence primer
 GCTTTCAACCTTACAACA
Pf dhfr codon 51, 51, 59
Forward primer TGAGGGGAAAAAAAATGAGGTT
Biotinylated reverse primer (HPLC purified) AAGGTTTAAATTTTTTTGGAATGC
Sequence primer
 AAAGGAGTATTACCATGGA
Pf dhfr codon 108
Biotinylated forward primer CAAAATGTTGTAGTTATGGGAAGA
Reverse primer TTTAAATTTTTTTGGAATGCTTTC
Sequence primer
 TGGAATGCTTTCCCAG
Pf dhfr codon 164
Biotinylated forward primer (HPLC purified) GTTTTACTTGGGAAATTAAATTACT
Reverse primer TCTGGAAAAAATACATCACATTC
Sequence primer
 ACAACGGAACCTCCTA
Pf dhps codon 436, 437
Forward primer ACGTGCTGTTCAAAGAATGTTT
Biotinylated reverse primer (HPLC purified) GTTTCGCATCACATTTAACAATT
Sequence primer
 ATAGATATAGGTGGAGAATC
Pf dhps Codon 540
Biotinylated forward primer (HPLC purified) GCATAAAAGAGGAAATCCACATAC
Reverse primer CGAGGTATTCCATTTAATACAAGA
Sequence primer
 ATTATCATAATTTGTTAGTT
Pf dhps codon 581
Forward primer ACCTCGTTATAGGATACTATTTGA
Biotinylated reverse primer (HPLC purified) TCCAATAAAAAGTGGATACTCATC
Sequence primer
 TTGATATTGGATTAGGATTT
Pf dhps codon 613
Biotinylated forward primer (HPLC purified) TCCACTTTTTATTGGATATTCAAG
Reverse primer CTGAAACATCCAATTGTGTGAT
Sequence primer
 CATTCATGCAATGGG
kelch13
K13_PCR_F CGGAGTGACCAAATCTGGA
K13_PCR_R GGGAATCTGGTGGTAACAGC
K13_N1_F GCAAGCTGCCATTCATTTG
K13_N1_R GCCTTGTTGAAAGAAGCAGA
K13_N2_F CGCCAGCATTGTTGACTAAT
K13_N2_R GCGGAGTAGTAGCGAGAAT

*Pyrosequencing using Pyromark PCR Kit protocol (QIAGEN, https://www.qiagen.com). cytb, cytochrome b; dhfr, dihydrofolate reductase; dhps, dihydropteroate synthetase; HPLC, high performance liquid chromatography; MDR, multidrug resistance; Pf, P. falciparum; pfcrt, P. falciparum chloroquine resistance transporter †See (10).

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