We describe a simple method for detection of
Plasmodium vivax and
Plasmodium falciparum infection in anophelines
using a triplex TaqMan real-time polymerase chain reaction (PCR) assay (18S rRNA). We tested the assay on
Anopheles darlingi
and
Anopheles stephensi
colony mosquitoes fed with
Plasmodium-infected blood meals and in
duplicate on field collected
An. darlingi. We compared the real-time PCR results of colony-infected and field collected
An. darlingi, separately, to a conventional PCR method. We determined that a cytochrome b-PCR method
was only 3.33% as sensitive and 93.38% as specific as our real-time PCR assay with field-collected samples. We
demonstrate that this assay is sensitive, specific and reproducible.