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Background  Malaria cases persist in some remote areas in Sabah and Sarawak despite the ongoing and largely successful malaria control programme conducted by the Vector Borne Disease Control Programme, Ministry Of Health, Malaysia. Point mutations in the genes that encode the two enzymes involved in the folate biosynthesis pathway, dihydrofolate reductase (DHFR) and dihydropteroate synthase (DHPS) enzymes confer resistance to pyrimethamine and sulfadoxine respectively, in both  Plasmodium falciparum  and  P. vivax . The aim of the current study was to determine the mutation on both  pvdhfr  at codon 13, 33, 57, 58, 61, 117, and 173 and  pvdhps  genes at codon 383 and 553, which are potentially associated with resistance to pyrimethamine and sulfadoxine in  P. vivax  samples in Sabah.    Methods  Every individual was screened for presence of malaria infection using a commercial rapid dipstick assay, ParaMax-3™ (Zephyr Biomedical, India). Individuals tested positive for  P. vivax  had blood collected and parasite DNA extracted. The  pvdhfr  and  pvdhps  genes were amplified by nested-PCR. Restriction fragment length polymorphism (RFLP) was carried out for detection of specific mutations in  pvdhfr  at codons 13Leu, 33Leu, 57Ile/Leu, 58Arg, 61Met, 117Asn/Thr, and 173Leu and  pvdhps  at codons 383Gly and 553Gly. The PCR–RFLP products were analysed using the Agilent 2100 Bioanalyzer (Agilent Technology, AS).    Results  A total of 619 and 2119 individuals from Kalabakan and Kota Marudu, respectively participated in the study. In Kalabakan and Kota Marudu, 9.37 and 2.45 % were tested positive for malaria and the positivity for  P. vivax  infection was 4.2 and 0.52 %, respectively. No mutation was observed at codon 13, 33 and 173 on  pvdhfr  and at codon 553 on  pvdhps  gene on samples from Kalabakan and Kota Marudu. One-hundred per cent mutations on  pvdhfr  were at 57Leu and 117Thr. Mutation at 58Arg and 61Met was observed to be higher in Kota Marudu 72.73 %. Mutation at 383Gly on  pvdhps  was highest in Kalabakan with 80.77 %. There are four distinct haplotypes of  pvdhfr / pvdhps  combination.    Conclusions  The presence of triple and quintuple mutation combination suggest that the  P. vivax  isolates exhibit a high degree of resistant to sulfadoxine, pyrimethamine and sulfadoxine-pyrimethamine combination therapy.

Mutations of pvdhfr and pvdhps genes in vivax endemic-malaria areas in Kota Marudu and Kalabakan, Sabah