A multiplex PCR system for Taeniid infections in definitive and intermediate hosts

The family taeniidae includes several species of veterinary and public health importance, but diagnosis of the etiological agent in definitive and intermediate hosts often relies on labor intensive and less specific morphometric criteria. In the present study, a new multiplex PCR system, based on five primers targeting the 18S rDNA and ITS2 sequences, produced unequivocal banding pattern for a range of taeniids. Identification at the species level by the multiplex PCR was evaluated in comparison to classical morphological identification and sequencing. Compared to sequencing, the multiplex PCR identified Taenia to species in 36 out of 43 infections in carnivores (83.7%, 69 - 93; 95% CI), while only 20 cases were identified by morphology (46.5%, 31 - 62; 95% CI). Among rodent infections, the multiplex PCR identified Taenia to species in 85 out of 94 infections (90.4%, 83 – 96; 95% CI), whereas only 55 cases were identified by morphology (58.5%, 48 – 69; 95% CI). In foxes, T. polyacantha was more prevalent than T. crassiceps and T. taeniaeformis was detected in five cats, while T. martis and T. mustelae were identified in a stone marten and a pole cat, respectively. In rodents, T. mustelae and T. polyacantha were most prevalent in rural forests, whereas T. taeniaeformis was the dominant species in urban forests and the sole species found in domestic gardens. Analysis of sequence variation and a phylogenetic tree based on sequenced of cox1 gene were performed. The multiplex PCR system proved its superiority to the classical morphological detection and could be applied in epidemiological studies and clinical diagnosis.