Development of PCR/dot blot assay for specific detection and differentiation of taeniid cestode eggs in canids.
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.
Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan.
Maria Teresa
MT
Armua-Fernandez
Nariaki
N
Nonaka
Tatsuya
T
Sakurai
Seita
S
Nakamura
Bruno
B
Gottstein
Peter
P
Deplazes
Isaac G K
IG
Phiri
Ken
K
Katakura
Yuzaburo
Y
Oku
D000818:Animals; D001483:Base Sequence; D003362:Cost-Benefit Analysis; D017931:DNA Primers; D018509:DNA, Helminth / Q000235:genetics; D004285:Dogs; D004443:Echinococcosis / Q000175:diagnosis* / Q000453:epidemiology / Q000235:genetics / Q000662:veterinary*; D004446:Echinococcus / Q000235:genetics*; D005243:Feces / Q000469:parasitology; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000235:genetics; D010270:Parasite Egg Count / Q000379:methods* / Q000662:veterinary; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000662:veterinary; D013045:Species Specificity; D013621:Taenia / Q000235:genetics*; D015024:Zambia / Q000453:epidemiology
D017931:DNA Primers; D018509:DNA, Helminth; D015345:Oligonucleotide Probes