Determining Molecular Markers Associated with Drug ‎Resistance in DHFR and DHPS genes of Plasmodium ‎Falciparum from Gombe L.G.A. Gombe State, Nigeria

Background: Malaria Chemoprevention depends on synthetic drugs, but the parasite is continuously developing resistance to the antimalarial armament, hence a consequential need for surveillance studies on the sensitivity of the drugs is felt. Therefore, the aim of this paper was to determine the presence of biomarkers associated with drug sensitivity in DHFR and DHPS gene of Plasmodium falciparum.Methods: ۲۰۰ blood samples were collected using vein puncture technique and they were analysed using Microscopy, RDT and PCR. DNA was extracted using Quick-DNA™ Miniprep extraction kit. Purity and concentration of the DNA were determined using Nanodrop Spectrophotometer. ۵۷ samples were selected for molecular analysis. Nested PCR was used to amplify PFDHFR and PFDHPS genes; all PCR reactions were carried out in ۲۵ µL reaction mixture (۵ µL DNA template, ۱ µL Primer, ۶.۵µldistilled water and ۱۲.۵ µL Master mix). The PCR products were subjected to electrophoresis using ۲% agarose gel. The amplicons were purified, sequenced and subjected to BLAST software.Results: Mutations were recorded from A۱۶V ۰۵(۸.۷۷%), N۵۱I ۱۸(۳۱.۵۸%), C۵۹R ۰۳(۵.۲۶%), I۱۶۴L ۱۲(۲۱.۰۵) variants of DHFR gene, while in DHPS gene, mutations were recorded from K۵۴۰E ۶(۱۰.۵۲%) variant. Conclusion: Basic Biomarkers of resistance in DHFR and DHPS gene were recorded from Gombe. 

Keywords:

Gombe , resistance , DHFR , DHPS , Malaria , Plasmodium falciparum