The detection of pfcrt and pfmdr1 point mutations as molecular markers of chloroquine drug resistance, Pahang, Malaysia

Abstract

BackgroundMalaria is still a public health problem in Malaysia with chloroquine (CQ) being the first-line drug in the treatment policy of uncomplicated malaria. There is a scarcity in information about the magnitude of Plasmodium falciparum CQ resistance. This study aims to investigate the presence of single point mutations in the P. falciparum chloroquine-resistance transporter gene (pfcrt) at codons 76, 271, 326, 356 and 371 and in P. falciparum multi-drug resistance-1 gene (pfmdr1) at codons 86 and 1246, as molecular markers of CQ resistance.MethodsA total of 75 P. falciparum blood samples were collected from different districts of Pahang state, Malaysia. Single nucleotide polymorphisms in pfcrt gene (codons 76, 271, 326, 356 and 371) and pfmdr1 gene (codons 86 and 1246) were analysed by using mutation-specific nested PCR and restriction fragment length polymorphism (PCR-RFLP) methods.ResultsMutations of pfcrt K76T and pfcrt R371I were the most prevalent among pfcrt gene mutations reported by this study; 52% and 77%, respectively. Other codons of the pfcrt gene and the positions 86 and 1246 of the pfmdr1 gene were found mostly of wild type. Significant associations of pfcrt K76T, pfcrt N326S and pfcrt I356T mutations with parasitaemia were also reported.ConclusionThe high existence of mutant pfcrt T76 may indicate the low susceptibility of P. falciparum isolates to CQ in Peninsular Malaysia. The findings of this study establish baseline data on the molecular markers of P. falciparum CQ resistance, which may help in the surveillance of drug resistance in Peninsular Malaysia.

Correlation of Molecular Markers, Pfmdr1-N86Y and Pfcrt-K76T, with In Vitro Chloroquine Resistant Plasmodium falciparum, Isolated in the Malaria Endemic States of Assam and Arunachal Pradesh, Northeast India

“…These findings corroborate well with the previous findings [12], [17], [19], [32]. In the present study, all the CQ resistant isolates showed mutational change at position 86Y confirming the role of this mutation in the CQ resistance.…”

Survey of chloroquine-resistant mutations in the Plasmodium falciparum pfcrt and pfmdr-1 genes in Hadhramout, Yemen

“…Similar findings have been reported from Malaysia and Iran (Rastaghi et al, 2008;Atroosh et al, 2012). No significant difference in the prevalence of pfcrt and pfmdr1 mutations between subclinical and clinical infection of malaria parasites was noted in this survey.…”

Prevalence of Plasmodium falciparum Molecular Markers of Antimalarial Drug Resistance in a Residual Malaria Focus Area in Sabah, Malaysia

“…falciparum CQ, PYR and SDX resistant previously reported in Pahang [9], Lundu, Sarawak [8] and Sabah: Kalabakan (Tawau Division) [6, 30], Keningau & Nabawan (Interior Division) [39], Kota Kinabalu (West Coast Division) [35], a Kota Marudu—present study, b Kudat [35] (Kudat Division). This map was generated by using SimpleMappr online software [40].…”

“…A hundred percent prevalence of K76T mutation was reported in Lundu, Sarawak in a study conducted in 1999 and 2000 (Fig 1 and Table 3) [8]. A previous study by Atroosh et al[9] have reported high prevalence rate of pfcrt K76T mutation in P . falciparum isolates in the Peninsular Malaysia (West Malaysia) area such as Pahang (Fig 1 and Table 3).…”

“…The widespread of resistance towards chloroquine (CQ) and sulphadoxine-pyrimethamine (SP) [4–9] has led Malaysia to change their antimalarial treatment policies to Riamet, a combination drug of artemether and lumefantrine (AL) for non-complicated P . falciparum malaria; while doxycycline are be given orally together with intravenous artesunate for complicated falciparum malaria treatment [2, 10].…”