title: Comparison of different commercial DNA extraction kits to detect Toxoplasma gondii oocysts in cat faeces.
abstract: The cat is the definitive host of Toxoplasma gondii and plays an important role in the transmission of this and other coccidian parasites, e.g. Hammondia hammondi, a protozoon closely related and morphologically similar to T. gondii. A number of techniques to detect T. gondii nucleic acids in feline faeces are described and several extraction kits for isolating pathogen DNA from faeces or soil are commercially available. To compare the performance of such kits with regard to isolating oocyst DNA, a feline sample that had tested negative for coccidian parasites including T. gondii and H. hammondi was spiked with 10(4), 10(3), 10(2), 50 and 10 H. hammondi oocysts. Several ready-to-use stool or soil kits and an in-house method were then used to extract parasite DNA from these spiked faecal samples. Of six kits tested, two were found suitable for the detection of H. hammondi oocysts DNA by the polymerase chain reaction (PCR) in faecal samples with a detection limit of 250 oocysts per 1 g of faecal sample. These two kits revealed a similar, even slightly lower detection limit (50 oocysts per 1 g of sample) when tested with faecal samples spiked with T gondii oocysts.
authors.affiliation: Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Epidemiology, Wusterhausen, Germany. daland.herrmann@fli.bund.de
authors.forename: Daland C
authors.initials: DC
authors.lastname: Herrmann
authors.forename: Aline
authors.initials: A
authors.lastname: Maksimov
authors.forename: Nikola
authors.initials: N
authors.lastname: Pantchev
authors.forename: Majda Globokar
authors.initials: MG
authors.lastname: Vrhovec
authors.forename: Franz J
authors.initials: FJ
authors.lastname: Conraths
authors.forename: Gereon
authors.initials: G
authors.lastname: Schares
mesh_terms: D000818:Animals; D002371:Cat Diseases / Q000175:diagnosis*; D002415:Cats; D005243:Feces / Q000469:parasitology*; D057230:Limit of Detection; D033661:Oocysts / Q000502:physiology*; D057089:Pathology, Molecular / Q000379:methods* / Q000592:standards; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D014122:Toxoplasma / Q000235:genetics / Q000302:isolation & purification*; D014124:Toxoplasmosis, Animal / Q000175:diagnosis*