Identification of taeniid eggs in the faeces from carnivores based on multiplex PCR using targets in mitochondrial DNA

• Published in: Parasitology Vol. 134; no. 6; pp. 911 - 920
• Main Authors: TRACHSEL, D., DEPLAZES, P., MATHIS, A.
• Format: Journal Article
• Language: English
• Published: Cambridge, UK Cambridge University Press 01.06.2007
• Subjects: Animals; Biological and medical sciences; Carnivora - parasitology; Carnivores; Cats; Cestoda - classification; Cestoda - genetics; cestode infections; Cestode Infections - parasitology; Cestode Infections - veterinary; DNA primers; DNA Primers - chemistry; DNA, Mitochondrial - genetics; Dogs; E. multilocularis; Echinococcus granulosus; Echinococcus multilocularis; Eggs; faecal samples; feces; Feces - parasitology; Flotation; Fundamental and applied biological sciences. Psychology; General aspects; General aspects and techniques. Study of several systematic groups. Models; Genotypes; Invertebrates; Mitochondrial DNA; mitochondrial genes; multiplex polymerase chain reaction; multiplex polymerase chain reaction (PCR); nucleotide sequences; ova; Parasite Egg Count - veterinary; parasite identification; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; quantitative PCR; Reproducibility of Results; restriction fragment length polymorphism; Sensitivity and Specificity; sequence analysis; Taenia; Taenia spp; tapeworms; quantitative PCR; faecal samples; genotypes; multiplex polymerase chain reaction (PCR); Taenia spp; Echinococcus granulosus; E. multilocularis; mitochondrial genes; Egg; Plathelmintha; Parasite; Genotype; Mitochondrial DNA; Cestoda; Polymerase chain reaction; Mitochondria; Taenia; Gene; Helmintha; Feces; Invertebrata
• ISSN: 0031-1820; 1469-8161
• DOI: 10.1017/S0031182007002235

A multiplex polymerase chain reaction (PCR) was evaluated for the identification of morphologically indistinguishable eggs of the taeniid tapeworms from carnivores using primers targeting mitochondrial genes. The primers for Echinococcus multilocularis (amplicon size 395 bp) were species-specific as assessed by in silico analysis and in the PCR using well-defined control samples. The design of primers that specifically amplify DNA from E. granulosus or Taenia spp. was not possible. The primers designed for E. granulosus also amplified DNA (117 bp) from E. vogeli, and those designed for Taenia spp. amplified products (267 bp) from species of Mesocestoides, Dipylidium and Diphyllobothrium. Nevertheless, as our diagnostic approach includes the concentration of taeniid eggs by sequential sieving and flotation, followed by their morphological detection, this non-specificity has limited practical importance. Sequence analysis of the corresponding amplicon can identify most of the described E. granulosus genotypes. Taenia spp. can be identified by direct sequencing of the 267 bp amplicon, or, for most species, by restriction fragment length polymorphism (RFLP) analysis. The multiplex PCR was readily able to detect 1 egg (estimated to contain 7000 targets, as determined by quantitative PCR). Having been validated using a panel of well-defined samples from carnivores with known infection status, this approach proved to be useful for the identification of taeniid eggs from both individual animals and for epidemiological studies.