Sequencing of PCR amplified fragment and sequence analysis
One of the PCR amplified fragments, generated from a macaque from Laos with P. knowlesi-specific primers, was sequenced using the Sanger dideoxy sequencing method [33] and analyzed on the ABI 3730 Capillary Electrophoresis Genetic Analyzer. The DNA sequences were aligned by Clustal W and analysed using DNASTAR software (Lasergene, USA) and DnaSP version 4.10.8. The phylogenetic analysis was inferred by the Neighbor-joining method [34] with Kimura 2-parameter, including transitions and transversions, using MEGA version 3.1. The reliability of the internal nodes of the tree was assessed by the bootstrap method after 1000 replicates.