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

Published online by Cambridge University Press:  09 February 2007

D. TRACHSEL ,
P. DEPLAZES  and
A. MATHIS
D. TRACHSEL
Affiliation:
Institute of Parasitology, Vetsuisse and Medical Faculty, University of Zurich, Winterthurerstrasse 266a, CH-8057 Zurich, Switzerland
P. DEPLAZES
Affiliation:
Institute of Parasitology, Vetsuisse and Medical Faculty, University of Zurich, Winterthurerstrasse 266a, CH-8057 Zurich, Switzerland
A. MATHIS*
Affiliation:
Institute of Parasitology, Vetsuisse and Medical Faculty, University of Zurich, Winterthurerstrasse 266a, CH-8057 Zurich, Switzerland
*
*Corresponding author. Tel: +41 (0)44 6358536. Fax: +41 (0)44 6358907. E-mail: alexander.mathis@access.unizh.ch
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Summary

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.

Keywords

Echinococcus granulosusE. multilocularisTaenia sppmultiplex polymerase chain reaction (PCR)genotypesfaecal samplesquantitative PCRmitochondrial genes

Information

Type
Research Article
Information
Parasitology , Volume 134 , Issue 6 , June 2007 , pp. 911 - 920
Copyright
Copyright © Cambridge University Press 2007

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