Original Research Article
Year: 2018 | Month: January | Volume: 8 | Issue: 1 | Pages: 36-42
Detection of Pfcrt Point Mutation as a Molecular Marker of Chloroquine Resistance in Plasmodium Falciparum, North India
Neha Kaushal, Haris M. Khan, Samia Kirmani, Adil Raza
Department of Microbiology, JNMC, AMU, Aligarh, UP, India
Corresponding Author: Neha Kaushal
Introduction: The development of chloroquine as an antimalarial drug and the subsequent evolution of drug-resistant Plasmodium strains has major impacts on global public health. In P. falciparum, chloroquine resistance is linked to multiple mutations in pfcrt (Plasmodium falciparum chloroquine resistance transporter). In particular, specific K76T point mutation is considered to be highly related to chloroquine resistance.
Objective: To detect pfcrt gene mutation in Plasmodium falciparum by PCR followed by restriction fragment length polymorphism.
Methods: The study was conducted in the Department of Microbiology, J. N. Medical College Hospital, Aligarh over a period of one and half years from February, 2014 to September, 2015. Blood sample was collected from 65 falciparum malaria infected patients. DNA was isolated, and mutation was detected by mutation specific PCR to give a 264-basepair product corresponding to amino acid residues 32 to 119 of the pfcrt gene product. Digestion of this PCR product with restriction enzyme Apo I results in two fragments of 128 and 136 basepairs if the CQ-sensitive haplotype (CVMNK) is present.
Results: Out of 65 samples, pfcrt K76T mutation was observed in only 23 (35.4%) of clinical isolates investigated. On RFLP analysis, Apo I enzyme did not digest any of the 23 PCR products. Conclusion: PCR and RFLP techniques provide a simple and rapid method of detecting genes that may predict chloroquine resistance. Although the identification of the mutation in the pfcrt provides a significant indicator of CQR, further studies are needed to determine the polymorphisms in the genes.
Key words: Chloroquine resistance, Plasmodium falciparum, pfcrt mutation