This PCR-sequencing approach functioned across human Plasmodium species including P. knowlesi. The COX-I primers had 100% of identity and 100% coverage with at least 26 different Plasmodium species including parasites from lizards, birds, rodents and non-human primates, which may be relevant in assessing malaria transmission in particular settings (e.g. forest border areas). This evidence concerns the gene MT-CO1 and malaria.