A new duplex real-time PCR for simultaneous detection and differentiation of and .

and are pathogenic avian parasites that infect a wide range of bird species. The pathologic potential of is controversial, whereas causes disease in many avian species. Infections are often asymptomatic in doves and pigeons; thus, columbids are presumed to represent the natural hosts for trichomonads. The detection of and is based on direct microscopic observation or a conventional PCR assay. Microscopy is not very sensitive, and identification of the trichomonads at the genus or species level is not possible. Conventional PCR assays have been developed primarily for phylogenetic studies, which detect a wide range of spp. but do not allow their differentiation. We developed a duplex real-time PCR (rtPCR) assay for the simultaneous detection and differentiation of and . We found that the rtPCR assay detected 10 plasmid DNA copies of and as few as 10 plasmid DNA copies of