Molecular analysis of bio-makers of Chloroquine resistance in Plasmodium falciparum isolate from gombe local government area, Gombe State, Nigeria

Abstract

Chloroquine was one of the most cheapest and effective chemotherapeutic drugs for Plasmodium falciparum-malaria, but for a long, the drug has been officially withdrawn in almost all malaria-endemic countries including Nigeria, due to the development of resistance by the parasite. Withdrawal of the drug may make the drug regains its efficacy. Therefore, this study aimed to determine the presence of Biomarkers associated with chloroquine resistance from Gombe Local Government Area, Gombe State, Nigeria after its withdrawal in 2005. Twenty hundred blood samples were collected from consented study subjects and analysed using Microscopy, RDT and PCR. DNA was extracted using Quick-DNA™ Miniprep (No. D4069), Purity and Concentration of the DNA were determined using Nanodrop Spectrophotometer. 57 true positive samples were selected for molecular analysis. Nested PCR was used to amplify the required codon (C72S, M74I, K76T and N75E) position of PCRT the gene of P. falciparum. Both Primary and Secondary PCR was carried out. The PCR products were subjected to electrophoresis in 2% agarose and stained with ethidium bromide. The amplicons were purified and sequenced, after which the sequenced products were subjected to BLAST software. Single Nucleotide Polymorphism was recorded from C72S and K76T with a prevalence of 05(8.80%) and 46(80.70%) respectively. Confirmed biomarkers of Chloroquine resistance are still present in P. falciparum isolate from Gombe L.G.A.

Keywords

• pcrt
• sensitivity
• molecular makers
• single nucleotide polymorphism