Ecotope-Based Entomological Surveillance and Molecular Xenomonitoring of Multidrug Resistant Malaria Parasites in Anopheles Vectors

Figure 3

Phylogenetic relationship of P. vivax dihydrofolate reductase (DHFR) homolog. The multiple sequence alignment of all representative DHFR homologs of P. vivax MDR malaria parasite populations was carried out at both nucleotide and amino acid levels. The phylogenetic reconstruction of haplotypes, which was tested 1000 times with bootstrap method, was constructed based on the maximum parsimony method by using the MEGA ver. 5.22 [76]. DHFR haplotypes and haplogroups (A to G) of geographically prone P. vivax parasites conferring point mutations responsible for resistance against antifols and sulfones/sulfonamides are shown in supplementary File S4 (see Supplementary Material available online at http://dx.doi.org/10.1155/2014/969531). As retrieved from the GenBank genome database, all the nucleotide or amino acid sequences (accession numbers) of P. vivax DHFR homologs correspond to the infection sources of the geographically prone P. vivax isolates including Pvdibbt-1 (KC121333), Pvachtk-1 (KC121334), and Pvachtk-2 (KC121335). The GenBank files of these three isolates are shown in supplementary Files S1 to S3. The qualifier of the submitted sequences pertaining to country or geographic area and submission year was used in the phylogenetic analysis. The isolation sources analyzed include the majority of patient isolates of geographically prone P. vivax parasites and a very lesser extent by *mosquito isolates and **laboratory strain of P. vivax asexual blood stage. Using standard country codes (http://www.eurogofed.org/calendar/codes.htm), country sources are denoted as DJ: Djibouti, GF: French Guiana, ID: Indonesia, IN: India, IR: Iran, KM: Comoros, KR: South Korea, MG: Madagascar, MM: Myanmar, PG: Papua New Guinea, PH: Philippines, SR: Surinam, SV: El Salvador, TH: Thailand, TL: East Timor, VN: Vietnam, and VU: Vanuatu.