title: Mutation analysis in pfmdr1 and pfmrp1 as potential candidate genes for artemisinin resistance in Plasmodium falciparum clinical isolates 4years after implementation of artemisinin combination therapy in Iran.
abstract: The emergence and spread of Plasmodium falciparum resistant to the commonly used anti-malarial drugs is a major challenge in the control and elimination of malaria. The present study provides information on genetic analysis in multidrug resistance 1 (pfmdr1) (N86Y/Y184F/S1034C/N1042D/F1226Y/D1246Y) and multidrug resistance protein 1 (pfmrp1) (H191Y/S437A/I876V/F1390I/K1466R) genes that are probably associated with artemisinin as well as chloroquine resistance transporter (pfcrt) 76T in P. falciparum clinical isolates (N=200) exposed to artemisinin-based combination therapy (ACT) 4years after its adoption in Iran. Also, the copy number of pfmdr1 gene was screened for its association with pfmdr1 mutations to incriminate artemisinin resistance. By using nested PCR-RFLP and sequencing analysis, none of the samples had any mutation at codons 1034, 1042, 1226 and 1246 of pfmdr1, while 86Y and 184F mutations were detected in 46% and 2% of the examined samples, respectively. Also, no significant difference was identified among analyzed samples collected before (baseline, 2002-2005) and after adoption of ACT (2007-2010) (P>0.05). As with pfmrp1 gene, the mutations at positions 191Y (76.5%), 437A (69.5%), 876V (64.5%) and 1390I (17%) were detected and no samples displayed mutation at codon 1466R. In total, 42.5% of the examined isolates carried both pfmdr1 86Y and pfcrt 76T and none of the parasites simultaneously harbored pfcrt 76T, pfmdr1 86Y, 184F and pfmrp1 191Y, 437A, 876V, 1390I mutations. In addition, the copy number of pfmdr1 gene (N = 1) was similar as a sensitive isolate, 3D7, to artemisinin. In summary, none of the potential mutations associated with artemisinin and its derivatives resistance was significantly changed 4years after adoption of ACT in Iran.
authors.affiliation: Malaria and Vector Research Group (MVRG), Biotechnology Research Center (BRC), Pasteur Institute of Iran, Tehran 1316943551, Iran.
authors.forename: Sakineh
authors.initials: S
authors.lastname: Pirahmadi
authors.forename: Sedigheh
authors.initials: S
authors.lastname: Zakeri
authors.forename: Mandana
authors.initials: M
authors.lastname: Afsharpad
authors.forename: Navid Dinparast
authors.initials: ND
authors.lastname: Djadid
mesh_terms: D037621:Artemisinins / Q000494:pharmacology* / Q000627:therapeutic use; D004351:Drug Resistance / Q000235:genetics*; D018628:Gene Dosage; D006239:Haplotypes; D006801:Humans; D007492:Iran / Q000453:epidemiology; D016778:Malaria, Falciparum / Q000188:drug therapy / Q000453:epidemiology; D058889:Molecular Typing; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000235:genetics*; D009154:Mutation; D059645:Mutation Rate; D010963:Plasmodium falciparum / Q000187:drug effects* / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D037621:Artemisinins; D027425:ATP-Binding Cassette, Sub-Family C Proteins; C513498:Mdr1 protein, Plasmodium falciparum; C031327:artemisinin