Gene Sequence Polymorphisms Mutations in PFMDR-1 and PFCRT-O Genes of Plasmodium falciparum

Abstract

Malaria still causes severe infection in human being and responsible for high mortality in the world. Different approaches have been developed to monitor the extent of antimalarial drug resistance and to det. the biol. mechanisms by which the parasite has evaded the action of the drug. The aim of this study was to find the mutations (N86Y and K76T) in chloroquine drug resistance genes in Plasmodium falciparum from human blood. Total 22 pos. samples of Plasmodium falciparum were included in this study. Chloroquine drug sensitivity testing was performed using method as per WHO III plate (micro test). Nested PCR was done for detection of pfcrt-o (Plasmodium falciparum chloroquine resistance transporter-o) and pfmdr-1 (Plasmodium falciparum multidrug resistance-1) genes of P. falciparum. Gene sequencing was done using Sanger method to find the mutations (N86Y) Pfmdr-1 and Pfcrt-o (K76T) assocd. with chloroquine drug resistance. Out of 22 P. falciparum 15 (68.18%) samples were Chloroquine resistance by method similar to (micro test) WHO III plate method and nested PCR which were also showed N86Y mutation in Pfmdr-1 (Plasmodium falciparum multidrug resistant-1) gene, and same no. K76T mutation was seen in pfcrt-o gene of Plasmodium falciparum. Gene sequencing is highly sensitive and specific mol. method and it could be able to differentiate between wild type and mutant type gene hence we recommend this method for point mutation which is most useful for detection of exact point where mutation occurred. [on SciFinder(R)]