International Journal of Current Microbiology and Applied Sciences (IJCMAS)
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Original Research Articles                      Volume : 5, Issue:10, October, 2016

PRINT ISSN : 2319-7692
Online ISSN : 2319-7706
Issues : 12 per year
Publisher : Excellent Publishers
Email : /
Editor-in-chief: Dr.M.Prakash
Index Copernicus ICV 2018: 95.39
NAAS RATING 2020: 5.38

Int.J.Curr.Microbiol.App.Sci.2016.5(10): 451-458

Gene Sequence Polymorphisms Mutations in PFMDR-1 and PFCRT-O Genes of Plasmodium falciparum
Gurjeet Singh1*, Raksha Singh1, A.D. Urhekar2 and Kshitija Rane3
1Department of Microbiology, NC Medical College and Hospital, Israna,Panipat-132107, Haryana, India
2Department of Microbiology, MGM Medical College and Hospital, Kamothe,Navi Mumbai-410209, Maharashtra, India
3Central Research Laboratory, MGM Medical College and Hospital, Kamothe,Navi Mumbai-410209, Maharashtra, India
*Corresponding author

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 determine the biologic 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 positive 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) associated 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 number K76T mutation was seen in pfcrt-o gene of Plasmodium falciparum. Gene sequencing is highly sensitive and specific molecular 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.

Keywords: Malaria drug resistance testing, Plasmodium falciparum, polymerase chain reaction,Navi Mumbai,Gene sequencing, microtitre plate.

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Gurjeet Singh, Raksha Singh, A.D. Urhekar and Kshitija Rane. 2016. Gene Sequence Polymorphisms Mutations in PFMDR-1 and PFCRT-O Genes of Plasmodium falciparum.Int.J.Curr.Microbiol.App.Sci. 5(10): 451-458. doi:
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