Application of Real-Time PCR for Detection of pfcrt Single Nucleotide Polymorphisms in Plasmodium falciparum in Southeast Iran

The main objective of present study was to evaluate the efficiency of real-time PCR technique for detection of 72-76 pfcrt gene mutations. In this regard, by using hybridization probes technology on light cycler instrument, the point mutations have been successfully detected in 28 blood samples collected from falciparum malaria patients in Chabahar in Southeast Iran. Our data showed pfcrt K76T mutation is present in 99% of samples. Three samples (10.7%) showed deletion in amino acid located in position 75, Asparagine, one of them was located at CVMK and two at SVMT alleles. Sequencing analysis of these samples was the same as real-time PCR result. Five samples were found with multi-clonal population of P. falciparum, which identified by the presence of the two peaks simultaneously in melting curve analysis. Real time PCR is found a sophisticated technique that can distinguish these mutations reliably with acceptable speed, high accuracy, sensitivity and reproducibility.