Prevalence of Sulfadoxine???pyrimethamine resistance-associated mutations in dhfr and dhps gene of Plasmodium falciparum isolates collected in Pala west region of Chad

Abstract


Souleymane IM*, TOURE OA, Mbanga D, Clément KH, Berenger AA, Baba C, Denis MM, Ephraim D, Passiri T, Honoré D, Marie VYJ, Samir B, Pascal R, Mireille D, Joséph DA

Sulfadoxine-Pyrimethamine (SP) is the recommended drug for Intermittent Preventive Therapy (IPT) in pregnant women in Chad since 2005. There are few published reports of mutations in dihydropteroate synthetase (dhps) and dihydrofolate reductase (dhfr) genes in P.falciparum populations in Chad. This study was assessed to determine prevalence of dhfr and dhps polymorphisms in P. falciparum isolates collected in Pala west region of Chad. P. falciparum isolates were collected from children under five years with a confirmed diagnosis of malaria at Pala hospital in the western region of Chad. The parasite DNA was extracted from the blood sampled on Whatman® filter paper by the Qiagen DNA Mini Kit method (Qiagen, Valencia.CA) according to the manufacturer's instructions. dhfr and dhps gene were amplified using nested PCR and subsequently sequenced. A total of 30 isolates were sequenced and analyzed. The prevalence of the Asn-51-Ile, Cys-59-Arg and Ser-108-Asn mutations of the dhfr gene were 23.3 %, 30% and 33.3%, respectively. The mutations Ala-16-Val, Ile-16-Leu and Ser-108-Thr were not detected. Ser-436-Ala, Ala-437-Gly mutations of the dhps gene were found in 23.3% and 16.7%. Lys-540-Glu, Ala-581-Gly, Ala-613-Ser mutations and Phe-436 codon were not observed. The wild type dhfr and dhps isolates represented 53.3% and 66.7% of the analyzed sequences. The triple mutant dhfr and double mutant dhps were respectively 3.33% and 6.7%.This study provide baseline prevalence dhfr and dhps polymorphisms in Chad. Deployment of SP for IPTp should be encouraged in Chad, and the corresponding resistance markers be regularly monitored.

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