Development of PCR/dot blot assay for specific detection and differentiation of taeniid cestode eggs in canids

Armua-Fernandez, M.T.; Nonaka, N.; Sakurai, T.; Nakamura, S.; Gottstein, Bruno; Deplazes, P.; Phiri, I.G.; Katakura, K.; Oku, Y. (2011). Development of PCR/dot blot assay for specific detection and differentiation of taeniid cestode eggs in canids. Parasitology international, 60(1), pp. 84-9. Amsterdam: Elsevier 10.1016/j.parint.2010.11.005

Text 1-s2.0-S1383576910001790-main.pdf - Published Version Restricted to registered users only Available under License Publisher holds Copyright. Download (1MB)

We report the development of a colourimetric PCR/dot blot assay targeting the mitochondrial gene NADH dehydrogenase subunit 1 (nad1) for differential diagnosis of taeniid eggs. Partial sequences of the cestode nad1 gene were aligned and new primers were designed based on conserved regions. Species-specific oligonucleotide probes (S-SONP) for canine taeniid cestodes were then designed manually based on the variable region between the conserved primers. Specifically, S-SONP were designed for the Taenia crassiceps, T. hydatigena, T. multiceps, T. ovis, T. taeniaeformis, Echinococcus granulosus (genotype 1), E. multilocularis and E. vogeli. Each probe showed high specificity as no cross-hybridisation with any amplified nad1 fragment was observed. We evaluated the assay using 49 taeniid egg-positive samples collected from dogs in Zambia. DNA from 5 to 10 eggs was extracted in each sample. Using the PCR/dot blot assay, the probes successfully detected PCR products from T. hydatigena in 42 samples, T. multiceps in 3 samples, and both species (mixed infection) in the remaining 4 samples. The results indicate that the PCR/dot blot assay is a reliable alternative for differential diagnosis of taeniid eggs in faecal samples.

Interest & Impact

Downloads

1 since deposited on 04 Oct 2013
0 in the past 12 months

Citations

5 Citations in Web of Science ®
6 Citations in Scopus

Search

in Google Scholar™

Services

Actions (login required)

Edit item

Actions (login required)

Edit item