Molecular Detection and Identification of Plasmodium sp Isolated from Cynomolgus Monkeys in Bogor-Indonesia Upik Kesumawati Hadi1, Uus Saepuloh2, Lis Rosmanah2, Susi Soviana1, Huda Shalahudin Darusman1,2
1Faculty of Veterinary Medicine, IPB University, Jl. Agatis, Dramaga, Bogor,
16680, Indonesia
2Primate Research Center, IPB University, Jl. Lodaya II/5, Bogor, 16151,
Indonesia
Abstract
Background: Asian macaques are natural host a number of Plasmodium parasites species. Some of these monkey malaria parasites are able to infect humans and having the potential to causes zoonotic infections. This study was conducted to update the prevalence of monkey malaria parasites in Bogor-Indonesia based on molecular detection and identification, especially in cynomolgus monkeys which has a wide range of geographic distribution and shares extensive habitats with humans. The information is needed to evaluate the status of simian malaria among macaques in Bogor area and to study the potential hazard to human health. Therefore, this updated data will provide adequate information for the implementation of malaria control strategies in the future, as well as to develop the potential malaria vaccine using the monkey as animal model.
Methods: Blood samples of 274 cynomolgus monkeys (Macaca fascicularis) were collected and identified by microscopy. The positive blood samples were DNA extracted and analyzed using polymerase chain reaction (PCR) technique to amplify the small subunit ribosomal RNA (SSU rRNA) target gene using consensus primer for genus level of Plasmodium. The PCR positive samples then nucleotide sequenced in commercial sequencing services and the results were analyzed using BioEdit program and aligned using the BLAST program from NCBI. Phylogenetic trees were constructed to see the kinship of the Plasmodium using MEGA 6.0 with Neighbour-Joining (NJ) method. Bootstrapping was performed to assess the robustness of tree topologies by using 500 replicates.
Result: Thirty eight out of 274 samples that microscopy detected positive to Plasmodium sp were also positive by PCR technique resulted 1640 bp amplicon. Further analysis using nucleotide sequencing technique confirmed these positive samples were Plasmodium inui with more than 99% nucleotide identity compared to database in genbank. Phylogenetic tree analysis of SSU-rRNA partial gene showed that all o
Keywords: Plasmodium inui, small subunit ribosomal RNA, phylogenetic tree
