Quantitative Real-time PCR with TaqMan and SYBR Green for Estimation of Pfmdr1 Copy Number and Application for Monitoring of Antimalarial Drug Resistance

Abstract

Amplification of Plasmodium falciparum multi-drug resistant gene 1 (Pfmdr-1) has
been linked with resistance of antimalarial drugs, particularly mefloquine and artemisinin
derivatives. This could be used as a molecular marker for monitoring and surveillance of
antimalarial drug resistance. The objective of the study was to compare the correlation
between copy number of Pfmdr-1 gene analyzed by two different techniques, i.e., real-time
polymerase chain reaction (PCR) with Taqman and and real-time PCR with SYBR green
assay. Gene copy number was determined in of a total of 18 dried blood spot samples
collected onto filter paper from patients infected with P.falciparum malaria. DNA extraction
Kit was used to extract genomic P.falciparum DNA. The results showed good correlation of
the copy number analyzed by the two methods (Spearman correlation test, r = 0.852). Further
study should be performed with more number of P.falciparum isolates, together with clinical
(in vivo) response, to confirm the association between Pfmdr-1 amplification and resistance
to mefloquine-artemisinin combination.